Gene expression profiling of antidepressant action in the brain

ABSTRACT

Implementing gene expression to study drug action in the central nervous system is complicated by functional heterogeneity because of the existence of many different neuronal subtypes within the mammalian brain. The integration of laser capture microdissection (LCM) and RNA amplification with cDNA microarray technology allows for large-scale gene expression analysis at cellular level. Using this approach, we have generated gene expression profiles of imipramine, a reference antidepressant, and a new putative antidepressant, novelR1 in several laser-captured brain nuclei (locus coeruleus, dorsal raphe, hypothalamic paraventricular nucleus and hippocampus) of rats subjected to the chronic mild stress model (CMS) of depression.

CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of United States Provisional Application Ser. No. 60/238,374, filed on Oct. 6, 2000, and U.S. Provisional Application Ser. No. 60/295,782, filed on Jun. 4, 2001.

BACKGROUND OF THE INVENTION

[0002] Microarray technology represents a potentially powerful approach to identifying genes specifically expressed in different cell of tissue type [P. O. Brown and D. Botstein, “Exploring the new world of the genome with DNA Microarrays” (1999) Nat Genet, 21:33-7; D. Shalon, “Gene expression micro-arrays: a new tool for genomic research”, (1998) Pathol. Biol. (Paris), 46;107-9; S. M. Welford, J. Gregg, E. Chen, D. Garrison, P. H. Sorensen, C. T. Denny and S. F. Nelson, “Detection of differentially expressed genes in primary tumor tissues using representational differences analysis coupled to microarray hybridization” (1998) Nucleic Acids Res, 26:3059-65]. The application of microarray to the study of gene expression patterns in the central nervous system is complicated by functional heterogeneity because of the existence of many different neuronal subtypes within the mammalian brain [D. H. Geschwind, “Mice, microarrays, and the genetic diversity of the brain [In Process Citation]” (2000) Proc. Natl. Acad. Sci. USA, 97:10676-8; S. J. Watson, F. Meng, R. C. Thompson and H. Akil, “The ‘chip’ as a specific genetic tool” (2000) Biol Psychiatry, 48:1147-56]. This heterogeneity consists of literally hundreds of brain nuclei that are in close proximity to each other as well as many different neuronal subtypes in cortical layers of different brain regions. We have demonstrated in a previous study that the integration of laser capture microdissection (LCM), T7 based RNA amplification and cDNA microarrays allows to profile gene expression at cellular level [L. Luo, R. C. Salunga, H. Guo, A. Bittner, K. C. Joy, J. E. Galindo, H. Xiao, K. E. Rogers, J. Wan, M. R. Jackson and M. G. Erlander, “Gene expression profiles of laser-captured adjacent neuronal subtypes” (1999) [published erratum appears in Nat Med 1999 Mar; 5(3):355], Nat Med, 5:117-22.; R. C. Salunga, H. Guo, L. Luo, A. Bittner, K. C. Joy, J. Chambers, J. Wan, M. R. Jackson and M. G. Erlander, “Gene expression analysis via cDNA microarrays of laser capture microdissected cells from fixed tissue” (1999) In M. Schena (Ed.), “DNA microarrays a practical approach”, Oxford University Press, Oxford].

[0003] Recently, expression profiling using microarrays has been successfully used to study the action of the topical anesthetic dyclonine in S. cerevisiae [Hughes, 2000 #346]. Implementing gene expression to study drug action in the central nervous system is complicated by functional heterogeneity because of the existence of many different neuronal subtypes within the mammalian brain [Geschwind, 2000 #373] [Watson, 2000 #442]. This heterogeneity consists of literally hundreds of brain nuclei that are in close proximity to each other as well as many different neuronal subtypes in cortical layers of different brain regions. Therefore, we reasoned that to study the in-vivo action of drugs there needed to be from the technical side, the ability to (1) microdissect small numbers of neurons (˜100) from selected nuclei/subnuclei; (2) amplify the extracted mRNA in a reproducible manner; and (3) generate robust cDNA microarray data. The integration of laser capture microdissection (LCM), T7 based aRNA amplification and cDNA microarrays technology allows profiling of gene expression from specific cell type in situ [Luo, 1999 #352] [Salunga, 1999 #347].

[0004] Acute tricyclic antidepressants and other classes of antidepressant have a relatively well defined pharmacological action/target in-vitro of presynaptic reuptake inhibition of biogenic amines (norepinephrine, serotonin, dopamine). However, clinical efficacy requires three to six weeks suggesting that in-vivo a more complex mechanism occurs [Nestler, 1998 #409] [Stanford, 1996 #252]. Given this lag in efficacy, it is reasonable to assume that the mechanism of action of chronic antidepressant treatment in vivo may in part involve the modulation of gene expression. Therefore, gene expression profiling of selected brain nuclei may help to elucidate the chronic mechanisms of antidepressant action.

SUMMARY OF THE INVENTION

[0005] The present invention provides gene expression profiles at a cellular level of multiple brain nuclei after CMS±chronic treatment with antidepressant imipramine. Imipramine, a potent inhibitor of norepinephrine (NE) and serotonin (5-HT) uptake was selected as reference compound. In addition, a new putative antidepressant was examined to determine whether different in-vitro pharmacological properties but similar behavioral effects of imipramine and the new compound in the CMS model result in similar gene expression patterns. This new compound displays U2 adrenoceptor and 5-HT₇ receptor antagonism properties [Kennis, 2000 #394]. The present invention also provides potential new targets for drug discovery to identify compounds useful to treat depression and other neurological diseases and/or conditions.

BRIEF DESCRIPTION OF THE DRAWINGS

[0006]FIG. 1: Laser Capture Microdissection from Nissl-Stained Sections (10 μM).

[0007] Panel A: hippocampus including CA1, CA3 and DG before capture

[0008] Panel B: hippocampus after capture of CA1 cells (c) CA1 captured cells.

[0009] Panel C: Scale bar represent 200 μm.

[0010]FIG. 2: Body Weight and Sucrose Consumption in Animals Exposed to CMS.

[0011] Effect of chronic treatment with vehicle (1 ml/kg 2× per day), imipramine (10 mg/kg/day) and novelR1 (0.16 mg/kg, 2× per day)

[0012] Panel A: on the body weight, and

[0013] Panel B: the consumption of a 1% sucrose solution in controls (open symbols) and in animals exposed to CMS (closed symbols). Treatment commenced following 2 weeks of stress. Values are means +/−SEM.

[0014] *−p<0.05, **−p<0.01, *** −p<0.001; relative to vehicle- or drug-treated control groups.

[0015] −p<0.05, −p<0.01, −p<0.001; relative to drug-treated stressed animals at Week 0.

[0016]FIG. 3: Experimental Reproducibility of LCM, T7 Based aRNA Amplification and cDNA Microarray Procedure.

[0017] Pair-wise similarities of gene expression profiles measured by the linear correlation coefficient.

[0018] Panel A: average linear correlation coefficient (R) for each brain area (n =3 to 5 per group)

[0019] Panel B: examples of scatter plots showing the individual linear correlation coefficient for two groups of rats (control/vehicle DR and control/vehicle CA3).

[0020]FIG. 4: Gene Expression Profiling of Laser Captured Brain Nuclei of Vehicle/Control Rats.

[0021] Panel A: table showing the number of genes expressed in each the laser-captured nuclei (100 cells) according our criteria for a call of presence (i.e. 3 fold above background). The number of genes uniquely expressed in one of these nuclei is also given

[0022] Panel B: two-dimensional hierarchical cluster analysis of the 871 genes (horizontal axis) detected in at least one of the region investigated (vertical axis). Red, high intensity (log10 scale), black not expressed (below 3 fold the background). The 7 regions on the vertical axis were assembled into a dendrogram, where items are joined by very short branches if they are similar to each other and by increasingly longer branches as their similarity decreases

[0023] Panel C: distribution of several genes for specific laser-captured brain region, data represent the average fluorescence intensity. Note that the fluorescence intensity readings for vasopressin and oxytocin in PA and MG correspond to the maximal intensity.

[0024] * The anatomical distribution of these genes matched the distribution previously described in the literature: vasopressin [Hallbeck, 1999 #359], oxytocin [Burbach, 1987 #354], somatostatin [Fitzpatrick-McElligott, 1988 #358], dynorphin [Morris, 1986 #364], tyrosine hydroxylase [Tohyama, 1998 #348], serotonin transporter [Swan, 1997 #419], galanin [Tohyama, 1998 #348], RGS14 [Traver, 2000 #416], H₃ histamine receptor [Lovenberg, 1999 #338], mineralocorticoid receptor [Reul, 1985 #424].

[0025]FIG. 5: In situ Verification of the cDNA Microarray Data for Tyrosine Hydroxylase mRNA by ISHH.

[0026] Autoradiograms of frontal sections (left hemisphere) through the locus coeruleus hybridized with cRNA probes for tyrosine hydroxylase mRNA. (a, c, e) control; (b, d, f) stressed; (a, b) vehicle; (c, d) imipramine treated (10 mg/kg/day); (e, f) novelR1 treated (0.16 mg/kg, 2× per day) (g) Bound radioactivity of tyrosine hydroxylase cRNA probe in locus coeruleus of each group of animals. Values are means ±SEM n =3-5. Differences between control and CMS or drug treated groups were statistically significant. ***p<0.001. Scale bar represents 0.5 cm.

[0027] FIGS. 6: Overlapping and Non Overlapping Gene Expression Pattern in Unstressed and CMS Drug Treated Rats in the 7 Different Nuclei.

[0028] The diagrams show the number of genes with the indicated patterns.

[0029] Panel A: unstressed and CMS imipramine treated animals

[0030] Panel B: unstressed and CMS novelR1 treated animals

[0031] Panel C: uniquely changed in CMS imipramine and NOVELR1: the subset of genes with an accession number overlapping are listed.

DETAILED DESCRIPTION OF THE INVENTION

[0032] Gene expression profiling of antidepressants should be carried in the context of a behavioral model of depression. The chronic mild stress (CMS) model, a behavioral model of depression for which predictive, face and construct validities have been demonstrated was selected [Willner, 1992 #150] [Willner, 1997 #144] [Yadid, 2000 #422]. The CMS model was developed to mimic anhedonia, a core symptom of depressive disorders. Chronic sequential administration of a variety of mild stressors causes a decrease in responsiveness to rewards in rats, which is reversed by chronic administration of antidepressants drugs [Willner, 1992 #150]. The CMS model of depression also causes a decrease in body weight [Forbes, 1996 #413] but the magnitude of this decrease is minor and the validity, reliability and utility of the CMS is overall well recognized [Yadid, 2000 #422] [Argyropoulos, 1997 #205] #] [Willner, 1996 #226].

[0033] The locus coeruleus (LC), dorsal raphe (DR), hypothalamic paraventricular nucleus (PVN) magno and parvocellular division (MG, PA) and hippocampus (CA1, CA3 and DG) were selected for the present study. About one hundred cells were laser-captured in each of these nuclei. These regions were selected based on the location of the sites of action of the drugs and the critical neuroanatomical substrate for the stress response. In an autoradiographic analysis of the in vivo distribution of [³H]imipramine in rat brain, the locus coeruleus (LC) and the hippocampus have been shown to be the primary sites of action of imipramine [Duncan, 1991 #407]. Imipramine has also been shown to interfere with the serotoninergic transmission, therefore the dorsal raphe was selected in the present study. The LC is a critical neuroanatomical substrate for the stress response [Brady, 1994 #37] [Nestler, 1999 #240] [Valentino, 1998 #71], the PVN was added to the list of the investigated nuclei because of its role in the stress response [Weiss, 1994 #36]. Information on the neuroanatomical distribution of genes is critical for the process of understanding their function in the central nervous system. The integration of laser capture microdissection, T7 based RNA amplification and cDNA microarrays allows dissection of neuronal heterogeneity and at the same time to measure the gene expression level of thousands of genes. We have built a catalogue of rat brain-nuclei specific gene expression profiles. The following seven brain nuclei (100 cells) were analyzed: locus coeruleus (LC), dorsal raphe nucleus (DR), hypothalamic paraventricular (PVN, parvocellular division PA and magnocellular division MG) and hippocampus (CA1, CA3 and DG). Of the 2137 genes (unique mRNAs) interrogated, 871 (40.76%) gave a hybridization signal consistent with a call of presence in at least one brain region (i.e. at least three-fold above background level). The analysis was validated by the agreement of our findings with that of previously documented expression patterns of “signature” genes for specific brain nuclei. For example, abundant mRNA expression encoding the serotonin transporter was detected in DR cells but not detected in the other examined nuclei. The different nuclei investigated were found to have about four to seven percent in unique gene expression. The hierarchical clustering analysis shows that the two divisions of the PVN (PA and MG) were closely related and that similarly the three regions of the hippocampus (CA1, CA3 and DG) shared a similar gene expression profile. This study demonstrates the feasibility and utility of cellular brain nuclei profiling.

[0034] Using the LCM, T7 based amplification and cDNA microarray approach, we have generated a catalogue of cell specific gene expression profiles that can be used to determine which genes are expressed and where in the adult rat brain. Recently, Sandberg et al. [R. Sandberg, R. Yasuda, D. G. Pankratz, T. A. Carter, J. A. Del Rio, L. Wodicka, M. Mayford, D. J. Lockhart and C. Barlow, “Regional and strain-specific gene expression mapping in the adult mouse brain” (2000) Proc. Natl. Acad. Sci., 97:11038-43] and Zirlinger et al. [M. Zirlinger, G. Kreiman and D. J. Anderson, “Amygdala-enriched genes identified by microarray technology are restricted to specific amygdaloid subnuclei” (2001) Proc. Natl. Acad. Sci. 98:5270-5] have demonstrated the utility of brain region expression profiling. In these studies, gene expression analysis was performed on dissected brain regions. In the present study, a more neuroanatomically refined method (i.e. LCM) was introduced to isolate the cells of interest. The LCM technique allows the procurement of a selected cell population [S. J. Watson, F. Meng, R. C. Thompson and H. Akil, “The ‘chip’ as a specific genetic tool” (2000) Biol Psychiatry, 48:1147-56]. Brain tissues are composed of complex admixtures of different cell types, their biological meaningful analysis necessitate the procurement of a pure sample of the cells of interest. It is likely that general brain region dissection approaches would dilute the concentration of RNAs from neurons with RNAs from millions of potentially unrelated neurons and glia.

[0035] The correlation coefficient underlying the pair-wise similarities of gene expression profiles within one nuclei obtained in the present study demonstrate the reproducibility of the LCM, T7 RNA amplification and cDNA microarray approach. The high concordance between our data and published results validates the approach. However, some false negative findings were observed in the present study. Hallbeck et al. [M. Hallbeck, O. Hermanson and A. Blomqvist, “Distribution of preprovasopressin mRNA in the rat central nervous system” (1999) J. Comp. Neurol., 411:181-200] have reported the presence of low level of vasopressin mRNA in the hippocampus by in situ hybridization. In the present study, the level of vasopressin mRNA detected in the hippocampus was found between two- and three-fold different from the background and so, according to our criteria, was classified as not expressed. Low expression of somatostatin mRNA in the hippocampus has also been described in the literature [J. V. Priestley, M. Rethelyi and P. K. Lund, “Semi-quantitative analysis of somatostatin mRNA distribution in the rat central nervous system using in situ hybridization” (1991) J. Chem. Neuroanat., 4:131-53]. As observed for vasopressin, the intensity for somatostatin mRNA in the present study was below the criteria used for a call of presence. The stringent criteria used for a call of presence in the present study could account for these discrepancies. Sandberg et al. [R. Sandberg, R. Yasuda, D. G. Pankratz, T. A. Carter, J. A. Del Rio, L. Wodicka, M. Mayford, D. J. Lockhart and C. Barlow, “Regional and strain-specific gene expression mapping in the adult mouse brain” (2000) Proc. Natl. Acad. Sci., 97:11038-43] reported in their study that approximately 1% of the genes showing hybridization signals on the array were detected as differentially expressed between the four different brain regions investigated. In the present study, we found that the seven different nuclei investigated have about 4% to 7% in unique gene expression. The higher percentage of uniquely expressed found in our study could reflect the higher sensitivity achieved by the LCM approach.

[0036] In this study, we were able to classify the different laser-captured brain nuclei by hierarchical clustering analysis. As expected, we found that the two divisions of the PVN (PA and MG) were closely related and, similarly, the three regions of the hippocampus shared similar gene expression profiles. The dendrogram in FIG. 3 represents a new approach to classify brain nuclei. In conclusion, this study demonstrates the feasibility and the utility of cellular brain nuclei profiling. The following examples are intended to illustrate but not limit the present invention.

EXAMPLE 1

[0037] Behavioral Procedure

[0038] The stress procedure was conducted on male Wistar rats as previously described [Dziedzicka-Wasylewska, 1997 #224]. On the basis of sucrose intake scores following two weeks of stress, both stressed and control groups were divided further into matched subgroups (n=8), and for the subsequent five weeks received twice-daily intraperitoneal injections of vehicle (isotonic mannitol solution, pH 4.0, 1 ml/kg) or novelR1 (0.16 mg/kg). Once-daily intraperitoneal injections of imipramine (10 mg/kg) were used as the reference treatment. At this dose, imipramine has been shown to cause stable and comparable effects after chronic treatment in all previous studies using this experimental protocol [Willner, 1997 #144]. Sucrose tests were carried out twenty-four hours following the last drug injection (in the case of b.i.d. regime, the second injections on the day preceding the sucrose tests were omitted).

[0039] Drugs

[0040] The following agents were used: imipramine HCl (RBI, Natick, Mass.) and novelR1.

[0041] Statistics

[0042] The behavioral results obtained in this study were analyzed by multiple analysis of variance with three between-subjects factors (stress/control, drug treatments and successive sucrose tests). Fisher's LSD test was used for post-hoc comparisons of means.

[0043] Tissue Preparation

[0044] Twenty-four hours after the final injection of vehicle, imipramine or novelR1, five representative animals from each group were decapitated, and their brains were removed, placed in cryomolds, covered with Tissue-Tek tissue embedding medium (OTC) and snap-frozen in dry ice-cold 2 methyl butane (−60° C.). Ten μm-thick coronal sections in the mid-regions of the PVN (bregma—1.80 mm), hippocampus (bregma—3.30 mm), DR (bregma—1.78 mm) and LC (bregma—9.80 mm) were cut using a cryostat, mounted on non-coated, clear microscope slides and immediately frozen on a block of dry ice. The sections were stored at −70° C.

[0045] Laser Capture Microdissection

[0046] A quick Nissl (cresyl violet acetate) staining was used to identify the neurons as described [Salunga, 1999 #347]. The PixCell II System from Arcturus Engineering, Inc. (Mountain View, Calif.) was used for LCM. According to the manufacturer's protocol, one hundred neurons were captured in each region of interest of each animal (LC, DR, MG, PA, CA1, CA3 and DG). Example of laser-captured cells in CA1 region is shown in FIG. 1.

[0047] RNA Extraction and Amplification of LCM Samples

[0048] The total RNA was extracted from the individual LCM samples as described [Salunga, 1999 #347] with modifications as specified below. For total RNA extraction, a small volume (20 μl/LCM sample) of denaturing buffer (Micro RNA isolation kit, Stratagene: San Diego, Calif.) containing 2-mercaptoethanol (10 μl/ml, Sigma, St Louis, Mo.) and polyinosine (300 ng/LCM sample, Sigma: St Louis, Mo.) was used. The samples were incubated at 42° C. for ten minutes, and loaded onto a pre-rinsed Microcon-100 column (Millipore: Bedford, Mass.). After two rinses with 500 μl Rnase-free water, the columns were inverted and the samples collected by brief centrifugation. The samples were then processed for two rounds of RNA amplification as described [Salunga, 1999 #347] with modifications for the DNA and RNA purification steps. Quiaquick PCR purification kit (Qiagen: Valencia, Calif.) and RNAeasy Mini Kit (Qiagen: Valencia, Calif.) were used for purification steps. Polyinosine (100 ng) was added to each sample before purification of DNA or aRNA.

[0049] cDNA Microarray

[0050] A cDNA microarray containing 2147 cDNA clones (corresponds to 2137 unique mRNAs) was used in this study. cDNA clones were obtained commercially from Research Genetics (IMAGE consortium), Incyte Genomics as well as internal sources. All clones were verified by DNA sequencing. Each clone was printed as two independent spots on a given chip. A contact pin microarrayer (Generation III Array Spotter, Molecular Dynamics) was used to spot the clones in duplicate. Microarrays were hybridized and scanned with a confocal laser scanner (Array Scanner, Molecular Dynamics) with excitation at 532nm and collection through a 550 to 600nm band width filter, the PMT set to 630V. Image analysis was performed as previously described [Salunga, 1999 #347].

[0051] Statistical Analysis

[0052] Hybridization of each sample was performed on two identical microarrays. Each microarray was normalized at the 75 percentile (intensity of the 75 percentile was scaled to 100). Intensity of each clone on an individual microarray was then measured as an average of the intensities of the two corresponding spots, and the intensity of each clone in the sample was further determined as average of the intensities on the two identical chips. The pair-wise similarity of gene expression profiles within a group of same samples (same cell type under identical treatment) was examined by scatter-plot and the correlation coefficient was calculated. Samples having a correlation coefficient of R <0.89 with respect to the majority were treated as out-liers and were excluded for further analysis (at least three rats were included in each group). The intensity of each clone in one particular type of samples was subsequently further defined as the median of intensities of that clone in remaining qualified samples in that group. The resulting intensity matrix (N×M) (N: number of clones; M: number of sample types) was further normalized (spline normalization) so that the distribution of the intensities of all the clones for each individual sample type was fitted to a smooth cubic B-spline. The spline normalization could reduce abnormality in the distribution of the intensities in particular samples caused by saturation in image processing.

[0053] In addition to the 2147 clones, fifteen plant cDNA were printed on the same microarray. The intensities of these fifteen plant cDNAs were measured and processed in the same manner as other clones, and used as negative controls. The average intensity of the fifteen plant genes in all M types of samples was defined as background. Each clone was then classified as absent in a cell population if its medium intensity was below the three-fold of the background; or as present if its medium intensity was three-fold above the background.

[0054] The ratio (fold change) of gene expression between two samples was calculated after applying *i minimum threshold at the background of plant genes (intensity lower than background was re-set to background). The criteria used to select statistically significant fold changes in gene expression between two samples was at least a 1.8 fold change. Hierarchical clustering was generated by using Cluster software (Stanford).

[0055] Quantitative In Situ Hybridization Histochemistry.

[0056] In situ hybridization histochemistry was performed according to the procedures described [Bonaventure, 1998 #353] on adjacent tissue sections of the one used for LCM. The plasmid (pBluescript II SK, Stratagene, San Diego, Calif.) containing the cDNA coding for tyrosine hydroxylase (GB:M10244) was linearized and [³⁵S]UTP labeled cRNA probe was generated by in vitro transcription with T7 polymerase. Sections were hybridized at 50° C. with 1.106 c.p.m of [³⁵S]UTP labeled cRNA probe (80 μl per section). After high stringency washes, the sections were exposed to Kodak BiomaxMR film for sixteen hours. [¹⁴C] standard strips (American Radiolabeled chemicals Inc., St Louis, Missouri) were co-exposed to allow densitometry. Autoradiograms were quantified with the aid of an MCID-M4 3.0 Image Analysis system (Imaging Research, St. Catharines, Ontario, Canada). Optical densities in the LC were transformed into levels of bound radioactivity after calibration of the image analyzer with grey-values generated by co-exposed standards. The hybridization signal was expressed as μCi/g. The Student's t-test was used for comparison between the groups.

[0057] Body Weight is Unchanged by CMS or Drug Treatment.

[0058] At the start of the study (Baseline) all animals weighed approximately 320 grams and after the initial two weeks of stress (Week 0) the body weights of control and stressed animals were comparable (375 +/−6 and 372 +/−9 g, respectively). At the end of the treatment period (Week 5), body weights of vehicle-treated control and stressed animals were comparable [Group effect: F(1,14)=0.011; p=0.919]. As compared to vehicle-treated groups, neither imipramine nor novelR1 had any significant effect on body weights in controls [IMI: F(1,14)=1.764; NS, novelR1: F(1,14)=1.241; NS] and in stressed [IMI: F(1,14)=1.002; NS, novelR1: F(1,14)=0.518; NS] (FIG. 2a).

[0059] Sucrose Intake is Normalized in CMS Rats by Chronic Imipramine/NovelR1 Treatment.

[0060] In the final baseline test, all animals drank approximately 13 grams of sucrose solution. Following the initial two weeks of stress (Week 0), the intakes remained at similar level in controls but fell to approximately 8 grams in stressed animals, resulting in a significant Group effect [F(1,14) 11.159; p<0.01]. As shown in FIG. 2b, such a difference between control and stressed animals treated with vehicle persisted for the remainder of the experiment [F(1 1,84)=5.563; p<0.001] and at the end of treatment period, the sucrose consumption in stressed animals receiving vehicle was significantly lower than that measured in control group (p<0.001). As compared to chronic vehicle administration, imipramine did not change the sucrose intake in control animals [Treatment effect: F(1,84)=3.344; NS, Treatment×Weeks interaction: F(5,84)=0.313; NS]. However, in stressed animals, imipramine gradually increased the sucrose consumption, resulting in a significant Treatment effect [F(1,84)=61.277; p<0.001], Treatment×Weeks interaction [F(5,84)=4.117; p<0.01], and Treatment×Group interaction [F(1,168)=13.353; p<0.001] after three weeks of treatment (p<0.001). This effect was maintained thereafter and at the end of treatment period (Week 5), the amount of sucrose solution consumed by these animals was comparable to that of vehicle-treated controls (p=0.168) and significantly higher than that of vehicle-treated stressed animals (p<0.001). The novelR1 like imipramine had no significant effect on the consumption of sucrose solution in control animals [Treatment effect: F(1,84)=0.315; NS, Treatment×Weeks interaction: F(5,84)=0.599; NS]. However, like imipramine, novelR1 fully reversed in stressed animals the CMS-induced deficit in sucrose consumption resulting in a significant Treatment effect [F(1.84)=51.123; p<0.001], Treatment×Weeks interaction [F(5,84)=2.882; p<0.05], and Treatment×Group interaction [F(1,168)=14.238; p<0.001].

[0061] As compared to Week 0 scores, the increase of sucrose intake reached statistical significance after two weeks of treatment with novelR1 (p=0.043). This effects was maintained thereafter and at the end of treatment period, the intake in this group of stressed animals did not differ from those measured in vehicle-treated controls (p=0.501) and were significantly higher than those of vehicle-treated stressed animals (p=0.001).

[0062] The Greatest Number of Genes Changing Their Expression Level in Response to the CMS Paradigm in Untreated Rats was Found in LC.

[0063] A total of 80 genes (3.79 %) were found to change their expression level in response to the CMS paradigm in untreated animals (versus the control/vehicle group). The greatest number of genes changing their expression level was observed in LC (53 genes)>DG (10 genes)>PA (9 genes), CA1 (9 genes)>DR (2 genes), CA3 (2 genes), MG (1 gene) in Table 1 (see below). TABLE 1 FOLD ACC. NO GENE (CMS vs ctrl) NUCLEUS Catecholamine synthesis M10144 Tyrosine hydroxylase 1.85 LC Neuropeptides A1045437 Neuropeptide Y** 2.11 DG AA955779 Galanin 2.06 PA M10088 dynorphin** −2.52 DG RNU10071 CART −1.90 LC Receptors, ion channels AF052746 Chloride channel CaCC 3.27 LC M81830 Somatostatin R2 (SSR2) 2.77 LC AF027955 GPR27 R −1.92 LC Z30425 Nuclear receptor/orphan −1.90 DG MB67/CAR L14618 Calcitonin R −1.81 PA L14618 Calcitonin R** −1.80 LC GABA signaling L08493 GABA-A R alpha4 subunit 2.24 CA1 X54673 GAT1 −2.05 LC Glutamate signaling X94552 mGlu7 R −2.21 LC M81886 glutamate R 1* −2.03 CA1 M64752 glutamate R 1* −1.89 CAl HSU08107 NMDA R subunit 2.23 LC Calcium AA819346 Parvalbumin** 1.93 DG M31178 calbindin D28 1.87 CA1 M31178 calbindin D28 −2.21 LC AA819345 Parvalbumin −1.99 LC L10284 Calnexin −1.87 LC Signal transduction D83407 ZAKI-4* −1.80 DR Heat shock protein M86389 Hsp27 −1.85 PA Plasticity X51396 MAP1B −2.11 CA1 X60370 MAP1B −1.83 CA1 Proteasome D29956 ubiquitin isopeptidase 1.97 LC D17296 Polyubiquitin −2.01 LC W07641 ubiquit-like protein SMT3B −2.00 LC Transcription factor Y10926 RBP-L 2.16 LC Y07688 NfiX −2.38 LC AF057691 TCF-3 −1.83 DR W65013 similar to X07384 GLI −1.85 CA3 PROTEIN p53 U67885 p53 binding prot 1 −2.37 LC U28789 PACT −2.01 LC Others X58526 L1 element −2.39 LC X58526 L1 element −2.35 DG 100256 Sequence 7 from U.S. Pat. 2.38 LC No. 4900811** 100256 Sequence 7 from U.S. Pat. −1.93 PA No. 4900811

[0064] These genes were classified in functional families (i.e. catecholamine synthesis, neuropeptides, receptors or ion channels, GABA signaling, glutamate signaling, calcium, signal transduction, heat-shock protein, plasticity, proteasome, transcription factors and p53). The genes that could be classified in these functional families and with an accession number are listed in Table 2 (containing 39 genes). TABLE 2 Number of genes detected in each laser-captured nuclei (100 cells) according our criteria for a call of presence (i.e. three-fold above background). The number of genes uniquely expressed in one of these nuclei is also given. Detected Unique CA1 499 32 CA3 527 22 DG 527 24 LC 513 43 DR 510 38 MG 519 21 PA 520 25

[0065] The increase in expression level of tyrosine hydroxylase mRNA (GB: M10244) in the LC of CMS/vehicle rats was further confirmed by quantitative in situ hybridization (FIG. 5a and b).

[0066] For the glutamate receptor 1 (GB: M81886, M64752) and the microtubule associated protein MAPI (GB: X51396, X60370) two different clones (from different species) were spotted on the microarray and consistent changes in gene expression level were observed for both clones as shown in Table 1. MAP1B plays an important role in the extension of axons and dendrites.

[0067] The chloride channel CaCC (GB: AF052746) and the somatostatin receptor 2 (GB: M81830) mRNAs showed the biggest amplitudes of changes of their expression levels in response to CMS (i.e 3.37-fold and 2.77-fold, respectively).

[0068] Six genes (five with an accession number) were found to change their expression level in the CMS/vehicle group in two different laser-captured nuclei (i.e. calcitonin receptor GB: L14618 in PA and LC, parvalbumin GB: AA819346 in DG and LC, calbindin D28 GB: M31178 in CA1 and LC, LI element GB: X58526 in LC and DG and GB: 100256 in LC and PA) (Table 1). Noteworthy, two calcium-ion binding protein (parvalbumin and calbindin D28) were found among these six genes.

[0069] At the end of the behavioral experiment, two different endpoints in sucrose consumption levels were observed: 13 grams of sucrose intake including the following groups vehicle/control, imipramine/control, imipramine/CMS, novelR1/control, novelR1/CMS) and 8 grams of sucrose intake including the vehicle/CMS group (FIG. 2b). The differences in gene expression level between these two groups (i.e. 8 grams versus 13 grams) reflects either changes causal to the decrease in sucrose consumption level or changes in response/adaptation to the CMS paradigm. A total of 30 genes (1.4%) were found to change their expression level in the untreated CMS group uniquely (i.e. no change in any of the other comparisons performed). The largest set of genes changing their expression level after CMS exposure uniquely was observed in LC (26)>PA (2)>DR (1), CA1 (1). These genes are listed in Table 1. Interestingly, in the GABA signaling pathway the expression level of both GABA-A receptor and GAT-1 was changed by the CMS paradigm uniquely.

[0070] Overall Both Drugs Changed the Expression Level of a Larger Set of Genes Than the CMS Paradigm and NovelR1 Changed the Expression Level of More Genes Than Imipramine in Unstressed Rats.

[0071] Overall imipramine and novelR1 were found to change the expression level of more genes than the CMS paradigm in control animals (CMS: 80 genes, 3.79%; imipramine: 86 genes, 4%; novelR1: 306 genes, 14%). These changes in gene expression levels induced by imipramine or novelR1 in unstressed rats do not reflect a change in sucrose uptake. In all, the nuclei investigated (excepted CA3 and MG), chronic treatment with novelR1 changed the expression level of more genes than imipramine in control animals (FIG. 6).

[0072] For both drugs genes that changed their expression levels in more than one of the investigated nuclei were found (imipramine: 12 genes: Table 3, R10747: 42 genes: Table 4). Both drugs were found to increase tyrosine hydroxylase mRNA levels in LC (fold increase versus control/vehicle: imipramine, 2.27; novelR1, 3.37). This up-regulation of tyrosine hydroxylase mRNA level was confirmed by in situ hybridization (FIGS. 5c and 5 d). Noteworthy, the pterin-4a-carbinolamine dehydratase (DCoH or PCD, GB: AA901316) which catalyses the recycling of the cofactor biopterin [Depaepe, 2000 #439] shared a similar expression pattern in LC (fold increase versus vehicle/control: imipramine, 1.98; novelR1, 2.70). DcoH is requested in association with tyrosine hydroxylase for dopamine biosynthesis. Co-localization of DCoH and tyrosine hydroxylase in LC neurons has been previously demonstrated [Depaepe, 2000 #439] TABLE 3 IMINCMS ACC. NO. GENE NAME vs. IMI NUCLEI Apoptosis U67885 p53 binding prot 1 3.12 DG U59746 Bcl-w 2.00 CA1 AF022223 BAG-1 1.83 DR Catecholamine synthesis AA142680 dopamine beta-hydroxylase 1.95 LC A1044610 dopa decarboxylase 1.81 LC Energy production V01556 mitochondrial nenome 2.36 DG fragment V00680 mitochondrial genes coding 1.90 CA3 for 16S and 12S V00681 mitochondrial genes for 1.84 MG 16S V00681 mitochondrial genes for −2.85 DR 16S X14848 mitochondrial genome −2.32 CA1 V00681 mitochondrial genes for −2.07 CA1 16S V00681 mitochondrial genes for −2.04 PA 16S V01556 mitochondrial genome −1.99 DR fragment V00680 mitochondrial genes coding −1.93 CA1 for 16S and 12S GABA signaling L08495 GABA receptor alpha6 2.56 DG X54673 GAT1 2.24 CA1 X54673 GAT1 2.14 DG X14767 GABA-A receptor, beta 1 1.96 CA1 subunit X15376 GABA-A receptor, gamma 2.00 DG 2 subunit Glutamate signaling AA297071 Similar to N-methyl-D- 1.97 LC aspartate glutamate receptor U16129 Glutamate receptor (GluR4) 2.27 DG Growth factors M21571 platelet-derived growth 1.96 CA1 factor (PDGFA) A chain U66198 FGF13 1.91 CA1 U66198 FGF13 2.27 DG Heat shock protein M19645 BiP/GRP78/ER resident 2.18 LC HSP70 D13627 TCP-1 Hsp60, theta subunit 2.06 DR AA823192 heat-shock protein 40 2.03 LC X15187 HSP90 1.91 DR M11942 70 kd heat-shock-like 1.89 LC protein X63368 HSJ1 1.88 CA1 Neuropeptides AI045437 neuropeptide Y 2.56 LC AA818532 gamma-preprotachykinin A 1.99 DR M10088 Dynorphin 1.86 MG Plasticity M16228 GAP-43 2.59 DG X51396 MAP1B −4.65 DG Proteasome U44839 ubiquitin C-terminal 4.27 DG hydrolase D10699 ubiquitin carboxyl-terminal 2.17 LC hydrolase D00761 proteasome beta 1 subunit 2.09 DR U44839 ubiquitin C-terminal 1.93 LC hydrolase AA859300 proteasomes C2 component 1.90 DR W07641 ubiquit.-like protein −2.79 DG SMT3B (SUMO-1) X99585 ubiquit.-like protein −2.60 DG SMT3B (SUMO-1) M29859 proteasomes C2 component −2.34 DG D00763 proteasome alpha 4 subunit −2.31 DG X99585 ubiquit.-like protein −1.99 MG SMT3B (SUMO-1) AF079564 ubiquitin carboxy-terminal −1.87 DR hydrolase ubp41 U75362 ubiquitin carboxy terminal −1.84 DG hydrolase M29859 proteasomes C2 component −1.83 MG AA859229 ubiquitin conjugating −1.82 DG enzyme Receptors, ions channels Y07521 Kv3.1a = NGK2 potassium 2.94 DG channel; V-gated U33267 Human glycine receptor 2.60 DG beta subunit (GLRB) mRNA, complete cds H3 Histamine Rec 2.09 DR L14618 Calcitonin Rec 2.06 LC X66842 Serotonin 2B Rec 2.05 MG Z30425 Nuclear receptor/orphan 2.03 MG MB67/CAR AF064876 BCNG-1 hyperpolarization 2.03 DG NSC channel = HCN1 = HAC2 AF027955 GPR27 Rec 1.96 DG Y10148 NTR2 receptor 1.94 DG H3 Histamine Rec* 1.93 LC X97121 Neurotensin 2 Rec 1.90 DG M81253 Kv3.4 gene 1.86 LC AA002993 Potassium channel Kv3.4a 1.85 DR (Raw3), V-gated U07225 Purinergic (P2U) 2.88 DG AI058645 Kv4.3 potassium channel 2.60 DG X66842 Serotonin 2B Rec 2.23 DR X66842 Setotonin 2B Rec 1.92 PA Z30425 Nuclear receptor/orphan 1.88 DR MB67/CAR U07225 Purinerpic (P2U) 1.87 PA U12194 voltage-gated Na + channel 1.83 LC beta-1 sub (SCN1B) gene M21948 dihydropyridine-sens Ca 1.83 CA3 channel alpha-2 sub M24898 Nuclear receptor alpha1 1.81 DG for thyroid horm Signa transduction D83407 ZAKI-4 3.41 DG M38194 extracellular signal- 2.26 LC regulated kinase 1 M12673 guanine nucleotide-binding −4.12 DR protein G-s, alpha M29871 ras-related C3 botulinum −2.69 LC toxin substrate (rac) D14425 Calcineurin −2.24 DG M60156 Go alpha −1.89 DR Transcription factor AF057691 TCF-3 3.35 DG AF012923 Wig-1 2.22 LC AF065386 Jab1 2.14 DR W65013 Similar to X07384 GLI −3.24 DG PROTEIN X17163 c-jun −2.50 DG X12740 C-Jun of the transcription −1.85 DG factor AP-1 Others M25888 Lipophilin 7.94 DG I00256 Sequence 7 from U.S. Pat. 5.89 DG No. 4900811 D28875 Osreonectin 3.83 DG U49099 cis-Golgi p28 (p28) 2.14 DG D28875 Osteonectin 2.07 PA P11661 nadh-ubiquinone ox 1.89 DG I00256 Sequence 7 from U.S. Pat. 1.89 LC No. 4900811 Z26634 ankyrin brain variant, 2.63 PA ANKB M89646 ribosomal protein S24 2.32 DR P11661 nadh-ubiquinone ox 2.10 CA1 M89646 ribosom protein S24 2.10 DG M25888 lipophilin 2.09 CA1 M89646 ribosornal protein S24 −1.98 LC U49099 cis-Golgi p28 (p28) −1.94 LC Z26634 ankyrin brain variant, −1.90 DG ANKB

[0073] TABLE 4 R107474NCMS ACC. No. GENE NAME vs. R107474 NUCLEI Apoptosis U59746 Bcl-w 2.11 DR AF022223 BAG-1 2.05 LC U59746 Bcl-w 1.85 CA1 U67885 p53 binding prot 1 1.84 PA Calcium AA819345 parvalbumin 3.18 CA1 AA819345 Parvalbumin 2.23 DR M31178 calbindin D28 2.20 CA1 catecholamine synthesis AA142680 dopamine beta- 2.34 LC hydroxylase* energy production M29341 glyceraldehyde-3- 2.24 LC phosphate dehydrogenase X02231 glyceraldehyde-3- 2.09 LC phosphate dehydrogenase V01556 mitochondria genome 2.07 MG fragment M35826 mitochondrial NADH- 1.96 PA dehydrogenase (NDI) glutamate signaling U08107 N-methyl-D-aspartate 1.85 CA3 receptor subunit U08107 N-methyl-D-aspartate 1.81 DG receptor subunit M81886 glutamate rec type 1 −2.58 CA1 HBGR1) U08 107 N-methyl-D-aspartate −2.46 DR receptor subunit M64752 glutamate rec sub −2.35 CA1 (GluH1) Plasticity M16228 GAP-43 2.03 LC X60370 MAP1B −2.21 CA3 receptors, ions channels Y07521 Kv3.1a = NGK2 2.41 PA potassium channel; V- gated AA875142 5HT3 receptor mRNA 2.08 LC H3 Histamine Rec* 2.05 LC AJ225124 Hyperpolarization-act 1.82 LC cation channel HAC3 AA427267 SCN9a = PN1 Sodium 1.81 LC Channel alpha, V-gated M59980 K Channel −2.24 CA1 AA819622 thrombin receptor −2.08 DG L28175 Prostaglandin EP2 −1.99 LC U12194 voltage-gated Na+ −1.83 LC channel beta-1 sub- (SCN1B) gene signal transduction AI071860 RGS14 2.64 CA1 M23612 GTPase-activating 2.16 CA1 protein (GAP) M29870 ras-related C3 botulinum 2.01 DR toxin substrate (rac) transcription factors W65013 Similar to X07384 GLI −2.16 CA1 PROTEIN. J04509 JUN-D prot −2.10 DR AF062566 Sp1 −1.94 LC Others M30952 28S ribosomal RNA gene 2.92 PA fragment U20553 P57 2.18 PA M30952 28S ribosomal RNA gene 2.14 MG fragment M93056 monocyte/neutrophil 2.12 PA elastase inhibitor X92106 Bleomycin hydrolase 2.08 PA U20553 P57 1.98 DG X92106 Bleomycin hydrolase 1.87 CA3 M29548 elongation factor 1-alpha 1.83 DR (EF1A) M93056 monocyte/neutrophil 1.80 DG elastase inhibitor M29548 elongation factor 1- −1.98 CA1 alpha (EF1A) M89646 ribosomal protein S24 −1.94 CA1 M89646 ribosornal protein S24 −1.89 CA1 X95591 ClD protein −1.85 CA1 X95591 ClD protein −1.80 CA3

[0074] The largest set of genes changing their expression level after chronic treatment with novelR1 was found in DG. Among these genes, the primary site of action of novelR1, the a2c adrenoceptor mRNA (GB: M99376) level was found to be increased (fold difference versus control/vehicle: 3.62).

[0075] Overlaps in the gene expression patterns of imipramine and novelR1 in unstressed animals were observed in PA (thirteen genes)>LC (ten genes), DG (ten genes)>DR (five genes)>CA1 (four genes)>MG (two genes). Among these overlapping genes, six (three with an accession number) were found to change their expression levels in two different nuclei (heat-shock protein 27, GB: M86389 in PA and DR; Gs, GB: M12773 in MG and PA; 5-HT₂B receptor, GB: X66842 in DG and CA1).

[0076] 35% of the Genes Changing Their Expression Level in Response to the CMS Paradigm were Antagonized by Both Drugs Treatment.

[0077] Imipramine and novelR1 were found to antagonize the changes in gene expression level induced by the CMS paradigm of a similar number of genes (imipramine: 53 genes, novelR1: 55 genes). This corresponds to 68% to 69% of the changes induced by the CMS paradigm. For twenty-eight of these genes overlap in the gene expression profile of imipramine and novelR1 were observed.

[0078] The increase in expression level of tyrosine hydroxylase mRNA induced by the CMS paradigm was not antagonized by chronic imipramine or novelR1 treatment in LC. The level of tyrosine hydroxylase mRNA were found to be up-regulated in the imipramine or novelR1/CMS group (fold increase versus vehicle/control: imipramine, 3.12; novelR1, 4.41). The up-regulation of tyrosine hydroxylase mRNA level was also confirmed by in situ hybridization (FIGS. 5e and 5 f). The level of dopamine β hydroxylase (GB: AA142680) mRNA, another enzyme involved in the catecholamine biosynthesis, was also increased by both drugs in CMS rats in LC (fold increase versus vehicle/control: imipramine, 1.95; novelR1, 2.34). Similarly, DcoH mRNA level was increased (fold increase versus vehicle/control: imipramine, 2.30; novelR1, 2.67).

[0079] In CMS rats Both Drugs Changed the Expression Level of a Similar Number of Genes in LC, DR, CA1, CA3 and PA but with Minor Overlaps in Their Expression Profiles.

[0080] In LC, DR, CA1, CA3 and PA, imipramine changed the expression level of a larger set of genes in CMS rats than in unstressed rats (FIG. 6a), whereas novelR1 changed the expression level of a similar number of genes in CMS and unstressed rats (FIG. 6b). Administration of imipramine or novelR1 to CMS animals normalized the sucrose intake to the control level (FIG. 2b). The genes that change their expression level in the drug treated/CMS group but not in the drug treated/control group could reflect a drug +CMS interaction. These changes in gene expression are either causal to the change in sucrose intake from 8 grams to 13 grams or are responding to the new behavioral state. Both drugs yield a similar number of genes reflecting the drug/CMS interaction in LC, DR, CA1, CA3 and PA. In DG and MG, the imipramine/CMS interaction induced greatest changes in gene expression than the novelR1/CMS interaction.

[0081] Minor overlaps in genes expression patterns between imipramine and novelR1 in CMS rats were observed: LC (four genes)>CA1 (two genes)>DR (one gene), PA (one gene). The overlapping genes with an accession numbers are listed in FIG. 6. Imipramine changed the expression level of a total of 184 genes in CMS rats, whereas novelR1 changed the expression level of 91 genes. The genes with an accession number were classified in functional families and are listed in Table 2 (imipramine) and Table 3 (novelR1). Thirty genes were found to change their expression level in more than one nucleus for imipramine and twelve genes for novelR1.

EXAMPLE 2

[0082] 2.1. Tissue Preparation

[0083] Five adult male Wistar rats (423 g±22) that were used as control animals in a behavioral paradigm were decapitated, their brains were removed, placed in cryomolds, covered with Tissue-Tek tissue embedding medium (OTC) and snap-frozen in dry ice-cold 2 methyl butane (−60° C.). Ten tm-thick coronal sections in the mid-regions of the PVN (bregma—1.80 mm), hippocampus (bregma—3.30 mm), DR (bregma—1.78 mm) and LC (bregma—9.80 mm) were cut using a cryostat, mounted on non-coated, clear microscope slides and immediately frozen on a block of dry ice. The sections were stored at −70° C. All efforts were made to minimize animal suffering and reduce the number of animals used.

[0084] 2.2. Laser Capture Microdissection

[0085] A quick Nissl (cresyl violet acetate) staining was used to identify the neurons as described [R. C. Salunga, H. Guo, L. Luo, A. Bittner, K. C. Joy, J. Chambers, J. Wan, M. R. Jackson and M. G. Erlander, “Gene expression analysis via cDNA microarrays of laser capture microdissected cells from fixed tissue” (1999) In M. Schena (Ed.), “DNA microarrays a practical approach”, Oxford University Press, Oxford]. The PixCell II System from Arcturus Engineering, Inc. (Mountain View, Calif.) was used for LCM. According to the manufacturer's protocol, one hundred neurons were captured in each region of interest of each animal (LC, DR, MG, PA, CA1, CA3 and DG). Example of laser-captured cells in CA1 region is shown in FIG. 1.

[0086] 2.3. RNA Extraction and Amplification of LCM Samples

[0087] The total RNA was extracted from the individual LCM samples as described [R. C. Salunga, H. Guo, L. Luo, A. Bittner, K. C. Joy, J. Chambers, J. Wan, M. R. Jackson and M. G. Erlander, “Gene expression analysis via cDNA microarrays of laser capture microdissected cells from fixed tissue” (1999) In M. Schena (Ed.), “DNA microarrays a practical approach”, Oxford University Press, Oxford] with modifications as specified below. For total RNA extraction, a small volume (20 μl/LCM sample) of denaturing buffer (Micro RNA isolation kit, Stratagene: San Diego, Calif.) containing 2-mercaptoethanol (10 μl/ml, Sigma: St Louis, Mo.) and polyinosine (300 ng/LCM sample, Sigma: St Louis, Mo.) was used. The samples were incubated at 42° C. for ten minutes, and loaded onto a pre-rinsed Microcon-100 column (Millipore: Bedford, Mass.). After two rinses with 500 μl Rnase-free water, the columns were inverted and the samples collected by brief centrifugation. The samples were then processed for two rounds of RNA amplification as described [R. C. Salunga, H. Guo, L. Luo, A. Bittner, K. C. Joy, J. Chambers, J. Wan, M. R. Jackson and M. G. Erlander, “Gene expression analysis via cDNA microarrays of laser capture microdissected cells from fixed tissue” (1999) In M. Schena (Ed.), “DNA microarrays a practical approach”, Oxford University Press, Oxford] with modifications for the DNA and RNA purification steps. Quiaquick PCR purification kit (Qiagen: Valencia, California) and RNAeasy Mini Kit (Qiagen: Valencia, California) were used for purification steps. Polyinosine (100 ng) was added to each sample before purification of DNA or aRNA.

[0088] 2.4. cDNA Microarray

[0089] A cDNA microarray containing 2147 cDNA clones (corresponds to 2137 unique mRNAs) was used in this study. cDNA clones were obtained commercially from Research Genetics (IMAGE consortium), Incyte Genomics, as well as internal sources. All clones were verified by DNA sequencing. Each clone was printed as two independent spots on a given chip. A contact pin microarrayer (Generation III Array Spotter, Molecular Dynamics) was used to spot the clones in duplicate. Microarrays were hybridized and scanned with a confocal laser scanner (Array Scanner, Molecular Dynamics) with excitation at 532 nm and collection through a 550 to 600 nm band width filter, the PMT set to 630V. Image analysis performed as previously described [R. C. Salunga, H. Guo, L. Luo, A. Bittner, K. C. Joy, J. Chambers, J. Wan, M. R. Jackson and M. G. Erlander, “Gene expression analysis via cDNA microarrays of laser capture microdissected cells from fixed tissue” (1999) In M. Schena (Ed.), “DNA microarrays a practical approach”, Oxford University Press, Oxford].

[0090] 2.5. Statistical Analysis

[0091] Hybridization of each sample was performed on two identical microarrays. Each microarray was normalized at the 75 percentile (intensity of the 75 percentile was scaled to 100). Intensity of each clone on an individual microarray was then measured as an average of the intensities of the two corresponding spots, and the intensity of each clone in the sample was further determined as average of the intensities on the two identical chips. The pair-wise similarity of gene expression profiles within a same cell type was examined by scatter-plot and the correlation coefficient was calculated. Samples having a correlation coefficient of R<0.89 with respect to the majority were treated as out-liers and were excluded for further analysis. The intensity of each clone in one particular nucleus was subsequently further defined as the median of intensities of that clone in remaining qualified samples in that nuclei. The resulting intensity matrix (N×M) (N: number of clones; M: number of sample types) was further normalized (spline normalization) so that the distribution of the intensities of all the clones for each individual sample type was fitted to a smooth cubic B-spline. The spline normalization could reduce abnormality in the distribution of the intensities in particular samples caused by saturation in image processing.

[0092] In addition to the 2147 clones, fifteen plant cDNA were printed on the same microarray. The intensities of these fifteen plant cDNAs were measured and processed in the same manner as other clones, and used as negative controls. The average intensity of the fifteen plant genes in all M types of samples was defined as background. Each clone was then classified as absent in a cell population if its medium intensity was below the three-fold of the background; or as present if its medium intensity was three-fold above the background. Hierarchical clustering was generated by using Cluster software (Stanford).

[0093] Gene Expression Profiling of Brain Nuclei is Reproducible.

[0094] Gene expression profiles were measured from seven different laser-captured brain nuclei in six different groups of rats (i.e. control/vehicle, CMS/vehicle, control/imipramine, CMS/imipramine, control/ novelR1, CMS/novelR1). To estimate experimental reproducibility within a group of rats, the pair-wise similarities of gene expression profiles were compared by calculating a linear correlation coefficient (FIG. 3). The average linear correlations coefficients (R) within one region ranged from 0.955 to 0.978 (n =3 to 5 per group, FIG. 3a). For examples, the correlation value for the DR in control/vehicle animals ranged from 0.95 to 0.98 (FIG. 3b). For CA3 the R values ranged from 0.96 to 0.98. These data suggest that the brain nuclei were clearly captured and that these nuclei are relatively homogenous in gene expression.

[0095] Different Brain Nuclei Have about 4% to 7% in Unique Gene Expression.

[0096] To further validate the LCM/cDNA microarray approach, the gene expression patterns of the seven laser-captured brain nuclei were compared within the control/vehicle groups (FIG. 4). Of the 2137 genes (unique mRNAs) analyzed, 871 (40.76%) gave a hybridization signal consistent with a call of presence in at least one brain region according our criteria (i.e. at least three-fold above background level). The number of genes detected within one region ranged from 499 to 527 (FIG. 4a). The number of genes unique to one of the seven investigated brain nuclei, according our criteria for a call of presence, ranged from 21 to 43 (FIG. 4a). The highest number of unique genes (43) was found in LC. All 419 genes that changed expression levels between the different areas of the brain are listed in Table 5.

[0097] To compare the overall gene expression patterns of the different brain areas two-dimensional hierarchical clustering was performed (FIG. 4b). Hierarchical clustering analysis showed similarities in expression patterns between the two divisions of the PVN and between the three regions of the hippocampus (FIG. 4b). For example, the dendrogram on the vertical axis shows that the two divisions of the PVN (PA and MG) were joined by a very short branch reflecting their similarities compared to the other areas (FIG. 4b) and the three regions of the hippocampus (CA1, CA3 and DG) were also located on a common branch. The LC was located on a different branch of the dendrogram, suggesting that this nuclei has the greatest difference in gene expression among the seven nuclei that were examined. This finding is consistent with the LC containing the highest number of genes that are enriched (described above).

[0098] The analysis was further validated by the agreement of our findings with that of previously documented expression patterns for “signature” genes for specific brain nuclei (FIG. 4c). For example, in our study, abundant mRNA expression encoding vasopressin (GB:AI072073) and oxytocin (GB:M88355) were detected in PA and MG cells, but not detected in the other examined nuclei. This finding is in agreement with previous studies [Hallbeck, 1999 #359] [Burbach, 1987 #354] [Tohyama, 1998 #348]. Other striking examples included mRNAs encoding tyrosine hydroxylase (GB: M10244) and serotonin transporter (GB: AA819178) to the LC and DR respectively (FIG. 4c). A B 1 Name 2 neuropeptide 9174| UI-R-E1-fg-b-04-0-UI| AA955779| Rat galanin (a neuropeptide) mRNA, complete cds 3 neuropeptide 9168| UI-R-E1-fg-g-12-0-UI| AA955767| Rat mRNA encoding rat pre-prosomatostatin (isolated from a medullary thyroid carcinoma) 4 8723| UI-R-A1-ev-b-11-0-UI| AA926266| Rat VL30 element mRNA 5 8637| UI-R-E0-br-e-03-0-UI| AA866450| Rattus norwegicus interleukin-2 receptor beta chain (p70/75) mRNA, complete cds 6 8615| UI-R-A0-aj-e-08-0-UI| AA866381| Rat mRNA for hnRNP protein, partial 7 neuropeptide 8610| UI-R-A0-bm-a-06-0-UI| AA866347| Rat mRNA encoding rat pre-prosomatostatin (isolated from a medullary thyroid carcinoma) 8 8569| UI-R-A1-el-b-12-0-UI| AA926045| Similar to AF020760 H sapiens serine protease (Omi) mRNA 9 apoptosis 8540| UI-R-A1-es-c-03-0-UI| AA926004| Rattus norvegicus mRNA for 14-3-3 protein eta-subtype, complete cds 10 ion channel 81| 832845| AA427267| SCN9a=PN1 Sodium Channel alpha, V-gated 11 7888| UI-R-A1-eg-b-04-0-UI| AA924903| Similar to HUMKG1C Human mRNA for KIAA0045 gene, mRNA 12 ion channel 786| UI-R-EO-bu-g-03-O-UI| AA875142| Rat 5HT3 receptor mRNA, complete cds 13 7732| UI-R-A1-dz-c-12-0-UI| AA924656| Similar to AB005298 H sapiens BAI 2 mRNA 14 735| UI-R-E0-ck-e-08-0-UI| AA874900| Similar to AC004021 kelch protein 15 7233| UI-R-A1-dp-d-03-0-UI| AA901316| Rattus norvegicus DCoH gene 16 6980| UI-R-E0-dk-d-12-0-UI| AA900886| Similar to MMUBF Mouse ubf gene for transcription factor UBFmRNA 17 68| 775900| AA276133| CIC-2 Chloride Channel, V-gated 18 6611| UI-R-C0-hc-e-09-0-UI| AF020760| Omi 19 hsp 6565| UI-R-G0-UI-h-03-0-UI| L39370| Human Hsp27 20 6544| UI-R-C1-ke-c-01-0-UI| L28175| Prostaglandin EP2 21 6510| UI-R-E1-fu-h-10-0-UI| L11667| Human cyclophilin-40 22 6505| 2076313| L10284| Human calnexin 23 6457| 918411| D42055| hNEDD-4/KIAA0093, hect domain protein 24 644| UI-R-E0-cq-c-04-0-UI| AA875241| Similar to MMUBCM4GN M. musculus UBcM4 mRNAmRNA 25 hsp 6439| 2064663| D13627| TCP-1 Hsp60, theta subunit 26 6407| 749025| M11723| Blood coagulation factor XII 27 6406| 660902| M112331 Cathepsin D 28 6351| UI-R-C0-IG-e-10-0-UI| M24898| Nuclear receptor alpha1 for thyroid horm 29 hsp 6332| UI-R-AD0-we-g-01-0-UI| M196451 Human BiP/GRP78/ER resident HSP70 30 GPCR 6296| 7006963431 M818301 Somatostatin (55R2) 31 G protein signaling 6240| 2088210| M60156| Go alpha (G-protein) 32 6232| 1152703| M58051| FGFR3 33 GPCR 6204| 700311648| U07225| Purinergic (P2U) 34 6150| UI-R-AE0-xe-f-02-0-UI| M94335| Akt1/PKB/RAC 35 6118| UI-R-CO-gs-d-06-0-UI| U39738| PAK3 36 6068| UI-R-AA1-aab-c-10-0-UI| U20553| P57 37 60664| 583865| U20372| Calcium channel beta 3 (CCHB3); V-gat,aux 38 proteasome 6031| 478404| W07641| ubiquit.-like protein SMT3B (SUMO-1) 39 GPCR 5997| 701899983| U71092| GPCR37 (Orphan, public=GPR24) 40 5981| 1885026| U59167| Desmin 41 ion channel 5977| 700254854| U57352| ASIC2=MDEG=BNC1 brain Na chan; amil sens 42 5958| 947959| U47036| CCR5 43 GPCR 5957| UI-R-Y0-acc-e-12-0-UI| U469231 GPCR8 (Orphan, public) 44 hsp 5934| 464942| X63368| H. sapiens HSJ1 mRNA, DnaJ homolog 45 ion channel 5897| UI-R-Y0-aci-c-12-0-UI| X16476| Kv2.1=DRK1 potassium channel; V-gated 46 hsp 5889| UI-R-A0-ab-h-12-0-UI| X15187| Human gp96/GRP94, ER HSP90 47 5873| 1211032| X04434| IGF1R 48 5855| 701644254| Z30425| Nuclear receptor/orphan MB67/CAR 49 ion channel 5845| 493098| Y07521| Kv3.1a=NGK2 potassium channel; V-gated 50 transcription factor 5844| 944489| Y00864| SCFR/c-kit 51 5832| 677053| X99585| ubiquit.-like protein SMT3B (SUMO-1) 52 GPCR 5825| 701293873| X94552| Glutamate metabotropic (mGlu7) 53 5824| 2182578| X92106| Bleomycin hydrolase 54 5821| 700230143| X89576| Matrix metalloproteinase MT2-MMP 55 5814| 368357| X85237| splicing factor SF3a120 56 5809| 316104| X82153| Cathepsin K 57 hsp 5789| UI-R-C0-hc-d-08-0-UI| X74801| Human TCP-1 HSP60, gamma subunit 58 apoptosis 5763| 100400| 100400| 14-3-3 gamma 59 GPCR 5704| 701035392| 258| H3 Histamine Rec (GPCR97) 60 apoptosis 5694| 95332| 95332| 14-3-3 eta 61 5681| 112173| 112173| calcineurin A2 62 4716| 583498| AA142680| S50200 S50200 dopamine beta-hydroxylase [mice, Genomic/mRNA, 2274 nt]. Blastn P. = 4.6E-167 63 4715| 580798| AA139498| L41351 HUMPROS Homo sapiens prostasin mRNA, complete cds. Blastn P. = 2.5E-141 64 4699| 604709| AA161790| X05026 HSRHOB Human rho mRNA (clone 12). Blastn P. =1.5E-204 65 4262| 577610| AA426192| hUBC6 66 39517| UI-R-AG0-ww-b-07-0-UI| U63421| sulfonylurea rec (SUR1) 67 transcription factor 39504| UI-R-01-kw-b-11-0-UI| AF012923| Wig-1 68 394961UI-R-AD1-zn-f-05-0-UIIU438441cyclinD3gene 69 39495| UI-R-02p-nz-g-03-0-UI| U28789| PACT 70 polyamine synthesis 39494| UI-R-Y0-acv-f-12-0-UI| J04791| DCOR 71 39490| UI-R-C0-jh-b-10-0-UI| Y14296| BTEB-1 72 transcription factor 39485| UI-R-C0-is-f-03-0-UI| X17163| c-jun 73 39477| UI-R-E1-gf-e-12-0-UI| Y07701| cytosol alanyl aminopeptidase 74 39475| UI-R-C3-ti-d-08-0-UI| Y07836| helix-loop-helix prot 75 apoptosis 39461| UI-R-C0-jq-f-07-0-UI| AF022223| BAG-1 76 39460| UI-R-Y0-abk-h-03-0-UI| U77615| crx homeodomain prot 77 39456| UI-R-AE0-xm-h-07-0-UI| AFO31384| TWIK-related acid-sens K+ channel (TASK) 78 GPCR 39452| UI-R-C2-nh-f-03-0-UI| X51585| Adrenergic alpha1B Rec 79 39451| UI-R-C2-mq-h-08-0-UI| U23781| A1-d prot 80 39428| UI-R-E0-cd-a-02-0-UI| D29678| CDKS 81 39424| UI-R-C0-jh-c-05-0-UI| U28068| neuroD 82 GPCR 39418| UI-R-Y0-mj-a-06-0-UI| M58316| Adrenergic alpha2C Rec 83 39416| UI-R-E0-de-a-06-0-UI| U1326| methionyl aminopeptidase II 84 39413| UI-R-A1-eg-e-06-0-UI| M62867| MSY-1 85 39404| UI-R-C2p-qw-a-08-0-UI| M31178| calbindin D28 86 39402| UI-R-AF1-aaq-g-02-0-UI| D76432| EF1 delta 87 39401| UI-R-BT0-qa-h-12-0-UI| U64828| SRC-1 88 39394| UI-R-C3-ty-c-12-0-UI| X15052| NCAM-140 89 transcription factor 39392| UI-R-G0-uh-h-09-0-UI| AF062566| Sp1 90 proteasome 39370| UI-R-E0-bv-f-08-0-UI| D26598| proteasome beta 3 subunit 91 39365| UI-R-E0-bv-d-02-0-UI| D29011| proteasome catalytic subunit 3 92 39358| UI-R-C2p-nt-b-07-0-UI| X65227| NMZ1 93 GPCR 39356| UI-R-AA1-aaa-d-04-0-UI| U15280| Oxytocin Rec 94 GPCR 39351| UI-R-C0-jn-c-07-0-UI| X97121| Neurotensin 2 Rec 95 39334| UI-R-C2p-ns-f-01-0-UI| U88716| Nkx-32 homeodomain prot 96 proteasome 39323| UI-R-E0-cy-e-09-0-UI| D00761| proteasome beta 1 subunit 97 prtoeasome 39321| UI-R-AC1-xw-f-06-0-UI| AF079564| ubiquitin carboxy-terminal hydrolase ubp41 98 transcription factor 39311| UI-R-C1-lq-e-10-0-UI| J04509| JUN-D prot 99 39296| UI-R-C2p-rc-d-02-0-UI| U19597| Cdk4/Cdk6 inhib prot 100 39290| UI-R-C1-kc-d-10-0-UI| U41741| MMUSF (USF) 101 39281| UI-R-C1-kb-d-12-0-UI| AF005885| cyclin G2 102 transcription factor 39274| UI-R-C0-hv-a-09-0-UI| AF065386| Jab1 103 39265| UI-R-Y0-ach-h-07-0-UI| X91753| SEF2 104 polyamine synthesis 39260| UI-R-AC0-yp-f-08-0-UI| D10706| OAZ 105 39254| UI-R-C2p-oh-h-09-0-UI| D29956| UBPY ubiquitin isopeptidase (H. sapiens) 106 39253| UI-R-C2p-ro-d-11-0-UI| AF017182| Sox10 107 39252| UI-R-C0-il-c-03-0-UI| U42556| freac-1 gene 108 proteasome 39236| UI-R-C3-tr-c-02-0-UI| S71381| proteasome beta 4 subunit 109 39212| 101689| 101689| Mouse transcriptional regulator RPD3 homolo 110 39209| 80164| 80164| Rat heavy neurofilament polypeptide NF-H 111 39195| 95418| 95418| Rat non-muscle myosin alkali light chain 112 39192| 476493| M97200| krueppel-like factor 113 39191| 700295357| M92340| IL6B 114 39175| 80186| 80186 Novel 115 39149| 102883| 102883 Novel 116 39140| 701293538| M21948 dihydropyridine-sens Ca channel alpha-2 sub 117 39133| 111979| 111979| Novel 118 39132| 701900803| M81886| glutamate rec type I (HBGR1) 119 39112| 872080| M26745| IL6 120 39111| 100756| 100756| Novel 121 39109| 109519| 109519| Rat PKG binding protein and substrat 122 ion channel 39094| 700771408| L08494| GABA-A receptor 123 39092| 113323| 113323| Rat light molecular-weight neurofila 124 39091| 112184| 112184| Novel 125 39083| 113568| 113568| Mouse neurofilament protein (NF-L) gene, 3′ flank 126 39075| 100749| 100749| Mouse transcriptional regulator RPD3 homolo 127 39068| 112215| 112215| retrovirus-related leucine zipper protein p40 - ra 128 39062| 700773553| AJ225124| hyperpolarization-act cation channel HAC3 129 39058| 112959| 112959| Novel 130 39057| 571654| X61506| E46 for E46 prot 131 39055| 112214| 112214| S-100 beta subunit [rats, Genomic, 423 nt, segment 132 39053| 112025| 112025| Rat ID-A element in the S-100 beta-subunit gene. 133 39049| 101164| 101164| P46783 Human 40s ribosomal protein 134 39025| 1366563| X51405| carboxypeptidase H 135 38988| 700183273| M10244| tyrosine hydroxylase 136 38987| 1972427| D14425| calcineurin 137 38986| 111692| 111692| Novel 138 38973| 113308| 113308| Rat light molecular-weight neurofilament 139 38971| 109491| 109491| Human MUM2 gene 140 apoptosis 38967| 1211267| U67885| p53 binding prot 1 141 38960| 113236| 113236| Rat myelin proteolipid (PLP) 142 38956| 95821| 95821| P09896 Human 143 38955| 95395| 95395| Rat light molecular-weight neurofilament 144 38592| 80905| 80905| Novel 145 38946| 103913| 103913| Novel 146 38945| 95793| 95793| Rat cytoplasmic dynein heavy chain 147 38941| 111147| 111147| Mouse B2 repeat in the 3′ flank of protein 53 (p53 148 38933| 1448814| M25889| Mylein basic port 149 GPCR 38929| 700148588| X66842| Serotonin 2B Rec 150 38922| 41932| U12194| voltage-gated Na+ channel beta-1 sub (SON1B) gene 151 38921| 700280626| S76145| Na/Cl dep neurotransmitter transporter 152 38917| 1364912| U36393| TFEB 153 38916| 477909| U68384| Mrg1b 154 38910| 112963| 112963| Mouse mitochondrial genes coding for three transfe 155 38909| 101166| 101166| Mouse neurofilament protein (NF-L) gene, 3′ flank. 156 38898| 106216| 106216| Horse serpin mRNA, complete cds. 157 38894| 100750| 100750| Sequence 1 from patent U.S. Pat. No. 5457026. 158 38884| 112218| 112218| S-100 beta subunit [rats, Genomic, 423 nt, segment 159 38879| 112228| 112228| Mouse TGF-beta-inducible protein (TSC-36) m 160 38875| 84488| 84488| Novel 161 38865| 1297351| M26657| germinal peptidyl-dipeptidaseA 162 proteasome 38863| 633539| D00763| proteasome alpha 4 subunit 163 38856| 463999| U63858| U57939 164 38845| 84487| 84487 Novel 165 38838| 101073| 101073| Rat PAG608 gene 166 38833| 1210758| AF057691| TCF-3 167 38829| 108622| 108622| ribosomal protein S24 homolog {clone 2-6} [rats, r 168 ion channel 38824| 679093| AF052746| chloride channel CaCC 169 38822| 103933| 103933| Rat gene for testicular cell adhesio 170 38816| 113570| 113570| Novel 171 apoptosis 38808| 111644| 111644| Rat RNA-binding protein SiahBP mRNA, 172 GPCR 38793| 701472025| M64236| Neurokinin 1 Rec 173 GPCR 38789| 402354| AF0279551 GPR27 Rec 174 38787| 111636| 111636| Novel 175 38783| 1211735| Y10926| RBP-L 176 38779| 95446| 95446| Mouse brain mRNA. 177 38772| 1162568| Y076881 NfiX gene 178 38768| 101714| 1017141 Novel 179 38764| 101216| 101216| Bos taurus ribosomal protein 52 mRNA, partial cds. 180 38761| 100721| 100721| Novel 181 proteasome 38756| 100763| 100763| P11661 Rat nadh-ubiquinone ox 182 neuropeptide 38737| 701905916| M10088| dynorphin 183 G protein signaling 38729| 95348| 95348| GTP-binding protein rab10 - rat 184 38725| 102725| 102725| Novel 185 ion channel 38721| 700247271| L08495| GABA receptor alpha6 186 GPCR 38716| 701031976| X63574| Somatostatin 3 Rec 187 38714| 112210| 112210| Mouse LB9 gene, partial sequence. 188 38712| 669616| U89412| PML isoform 1 189 38711| 716499| U90724| X-Proaminopeptidase(eukaryote) 190 GPCR 38710| 700369402| L14618| Calcitonin Rec 191 38701| 1331673| M11597| CGRP alpha 192 38694| 112194| 112194| Novel 193 38676| 701896910| M36418| GLR1 194 GPCR 38675| 701901569| M99376| Adrenergic alpha2C Rec 195 38667| 111664| 111664| Human (clone hh18) protein tyrosine phospha 196 38664| 700133895| 038450| 954 Rec 197 38663| 701921173| M64752| glutamate rec sub (GluH1) 198 38653| 106117| 106117| Novel 199 38623| 113304| 113304| Rat light molecular-weight neurofilament 200 38621| 444406| 444406| Novel 201 38614| 111621| 111621| Novel 202 38605| 108620| 108620| Rat receptor for hyaluronan-mediated 203 38602| 101693| 101693| Mouse ribosomal protein (ke-3) gene, exons 204 38601| 100748| 100748| Mouse transciptional regulator RPD3 homolo 205 38594| 113748| 113748| Mouse necdin mRNA, complete cds. 206 ion channel 38582| 700773668| M81253| Kv3.4 gene 207 38577| 113515| 113515| Novel 208 38558| 66940| 66840| Novel 209 38546| 96780| 96780| Bos taurus clone CSSM015 microsatellite DNA sequen 210 38545| 101138| 101138| Rat ribosomal protein S24 211 38536| 111254| 111254| Human C14orf3 protein 212 38533| 475889| AF020198| PBX2 213 38532| 103937| 103937| Rat heavy neurofilament (NF-H) polypeptide, partia 214 38528| 112216| 112216| S-100 beta subunit [rats, Genomic, 423 nt, segment 215 38522| 463924| T67463| other peptidase (C1A) 216 38508| 66531| 66531| Novel 217 38504| 84486| 84486| Novel 218 38491| 108519| 108519| Rat gamma-smooth muscle isoactin pro 219 38487| 112021| 112021| Rat brain ID transcript, clone bBC2- 220 38469| 95406| 95406| Rat light molecular-weight neurofilament 221 38463| 113514| 113514| Novel 222 38417| 79183| 79183| Novel 223 38404| 1039810| AA297071| Similar to N-methyl-D-aspartate glutamate receptor 224 38402| 701289315| X99792| capacitative Ca entry channel 1 225 ion channel 38386| 700495245| L08493| GABA-A receptor 226 37466| 386514| W65013| Similar to X07384 GLI PROTEIN. 227 37464| 386559| W65005| Mouse cDNA clone IMAGE:386559 5′ 228 36393| 387565| A18932401 Similar to L14935 Mouse neural retina-specific leucine zipper protein. 229 ion channel 33| 426170| AA002993| Potassium channel Kv3.4a (Raw3), V-gated 230 2319| UI-R-A0-af-e-06-0-UI| AA817907| Similar to MUSCDPK mouse cyclin-dependent kinase homologue (p130PITSL) mRNA, complete cdsmRNA 231 21709| UI-R-AF0-ya-b-04-0-UI| 988064T6| LVENNOT03 INCYTE 232 21692| UI-R-AC1-xn-e-07-0-UI| 817852H1| OVARTUT01 INCYTE 233 21664| 903057| 580795H1| NMDA channel-like 234 DNA repair 21573| 1054114| 3764982H1| human homologue of yeast RAD23 235 DNA repair 21458| UI-R-E0-ci-a-06-0-UI| 2583919H1| human homologue of yeast RAD23 236 21418| 1348327| 2049923H1| LIVRFET02 U43892 g1 167981 Mouse ABC transporter-7 mRNA, partial cd gb103rod 83 -55 237 DNA repair 21304| 420048| 1676240T6| cell cycle checkpoint rad9 gene 238 21251| 975800| 1275842F6| Eph-Erk 239 ion channel 21230| 748734| 2658404T6| SK2 calcium-activ potassium chann homolo 240 20977| UI-R-02p-np-g-01-0-UI| 485639T6| retinoic acid hydroxylase hP450RAI 241 20971| 2182407| 3369279H1| LON; ATP-Dep Proteas LA 242 20768| 96782| 96782| M58405 R. norvegicus thymosin beta-10 gene, complete cds. 1.0e-72 243 20765| 95816| 95816 X51396 Mouse MAP1B mRNA for MAP1B microtubule-associated 6.5e-99 244 axon outgrowth 20763| 95791| 95791| M16228 Rat neuronal growth protein 43 (GAP-43) mRNA, comp 9.7e-187 245 20762| 95494| 95494| M93056 Human mononcyte/neutrophil elastase inhibitor mRNA 1.9e-117 246 20760| 95467| 95467| U43747 Human frataxin (FRDA) mRNA, complete cds. 1.9e-107 247 20759| 95416| 95416| S77858 non-muscle myosin alkali light chain [rats, Spragu 2.9e-92 248 hsp 20754| 95386| 95386| M11942 Rat 70 kd heat-shock-like protein mRNA, complete c 9.0e-108 249 20751| 95321| 95321| G27093 human STS SHGC-31976. 1.5e-91 250 20749| 95308| 95308| S82024 SCG10=neuron-specific growth-associated protein/st 4.4e-102 251 20748| 94433| 94433| Z92839 Caenorhabditis elegans cosmid T08D2, complete sequ 0.89 252 20743| 84483| 84483| AC003043 Homo sapiens chromosome 17, clone HRPC1067M6, comp 2.9e-31 253 energy production 20740| 83925| 83925| X02231 Rat mRNA for glyceraldehyde-3-phosphate-dehydrogen 7.9e-210 254 20739| 83922| 83922| 569874 C-FABP=cutaneous fatty acid-binding protein [rats, 6.0e-194 255 20738| 83912183912| L37086 Homo sapiens FK-506 binding protein (fkbp12.6) gen 1.9e-151 256 20726| 82552| 82552| D17296 Rat mRNA for polyubiquitin (ten completely repetit 5.4e-90 257 20724| 82519| 82519| M81225 Rat farnesyltransferase alpha subunit mRNA, comple2.7e-164 258 20720| 82475| 82475| V00681 R. norvegicus mitochondrial genes for 16S rRNA, tRN 8.6e-82 259 20716| 82210| 82210| AF014955 Homo sapiens TFAR19 mRNA, complete cds. 1.3e-70 260 20710| 82096| 82096| X14848 Rat (R. norvegicus) mitochondrial genome. 1.0e-99 261 2071| UI-R-AO-at-f-11-0-UI| AA818381| Similar to MMCKIT Mouse c-kit mRNAmRNA 262 axon outgroth 20709| 82094| 82094| X03369 Rat mRNA for beta-tubulin T beta15. 5.7e-105 263 20708| 82093| 82093| G15458 human STS SHGC-16876. 6.7e-51 264 20703| 82047| 82047| M71245 Rat prostatein C3 subunit gene, complete cds. 3.7e-36 265 proteasome 20694| 81776| 817761045249 Rat mRNA for proteasome activator rPA28 subunit al 1.4e-178 266 20693| 81773| 81773| X68273 M. musculus mRNA for macrosialin. 1.0e-134 267 20690| 81743| 81743| X05608 Human gene for neurofilament subunit NF-L. 5.0e-38 268 20674| 80860| 80860| X95591 M. musculus mRNA for C1D protein. 1.7e-77 269 20673| 80835| 80835| AC000083 Home sapiens Chromosome 22q11.2 Cosmid Clone 68a1 0.65 270 20672| 80832| 80832| U16686 Rattus norvegicus defensin RatNP-1 precursor mRNA, 2.9e-117 271 20669| 80637| 80637| X70369 R. norvegicus mRNA for pro alpha 1 collagen type II 2.9e-81 272 20667| 80178| 80178| M58405 R. norvegicus thymosin beta-b gene, complete cds. 6.1e-79 273 20665| 80157| 80157| D83407 ZAKI-4 mRNA in human skin fibroblast, complete cds 1.2e-59 274 20661| 80013| 80013| G27219 human STS SHGC-30611. 2.6e-60 275 20655| 79192| 79192| V00680 Rat mitochondrial genes coding for 16S and 12S rRN 1.7e-109 276 20654| 79184| 79184| J04517 Rat high molecular weight neurofilament (NF-H) pro 5.3e-141 277 20591| 66897| 66897| M80899 Human novel protein AHNAK mRNA, partial sequence. 1.3e-38 278 20590| 66886| 66886| U52828 Human Cri-du-chat region mRNA, clone NIBA2. 2.2e-75 279 20588| 66841| 66841| X51406 Rat mRNA for carboxypeptidase E (EC 3.4.17.10). 2.4e-167 280 20587| 66839| 66839| M15563 Human MHC class II HLA-DR-beta-1 pseudogene (DR7), 0.79 281 20583| 66706| 66706| M30952 Orangutan 28S ribosomal RNA gene fragment. 9.8e-151 282 20582| 66692| 66692| L81910 Homo sapiens (subclone 3_c10 from PAC H74) DNA seq 0.00048 283 20581| 66688| 66688| U49099 Rattus nervegicus cis-Golgi p28 (p28) mRNA, comle 6.8e-193 284 20568| 115932| 115932| M25638 Rat smallest neurofilament protein (NF-L) mRNA, pa 6.7e-157 285 20566| 115916| 115916| X62952 R. norvegicus mRNA for vimentin. 2.8e-167 286 20563| 113752| 113752| AL021308 Human DNA sequence from cosmid U246D9 on chromosom 0.93 287 20562| 113749| 113749| M80840 M. musculus necdin mRNA, complete cds. 1.7e-71 288 20557| 113571| 113571| D28875 Rat mRNA for osteonectin, complete cds. 2.2e-171 289 20550| 113529| 113529| M38194 Rat extracellular signal-regulated kinase 1 mRNA, 2.8e-203 290 20545| 113318| 113318| I00256 Sequence 7 from U.S. Pat. No. 4900811. 1.3e-191 291 20541| 113285| 113285| X76985 R. norvegicus mRNA for latexin. 4.3e-171 292 20540| 113284| 113284| M23984 Rat mitochondrial proton/phosphate symporter mRNA, 2.7e-174 293 ion channel 20536| 113244| 113244| X15466 Rat mRNA for GABA(A) receptor beta-1 subunit. 7.3e-172 294 20532| 113167| 113167| AC002299 Homo sapiens Chromosome 16 BAC clone CIT987-SKA-11 0.84 295 20530| 113155| 113155| U40933 Caenorhabditis elegans cosmid F20D12. 0.38 296 20529| 113063| 113063| AC000399 Genomic sequence from Mouse 9, complete sequence. 6.5e-11 297 20520| 112951| 112951| AC002121 Genomic sequence from Mouse 11, complete sequence. 0.00031 298 20512| 112160| 112160| M14512 Rat Na+, K+-ATPase alpha(+) isoform catalytic subun 1.5e-259 299 20505| 111646| 111646| D28875 Rat mRNA for osteonectin, complete cds. 2.5e-206 300 20501| 111616| 111616| X58526| Mouse DNA for an L1 element. 1.3e-70 301 20500| 111613| 111613| M25888 Rat lipophilin mRNA, 3′ end. 4.4e-125 302 20496| 111441| 111441| U25844 Mus musculus serine proteinase inhibitor (SPI3) mR 1.6e-134 303 20476| 109494| 109494| M38188 Human unknown protein from clone pHGR74 mRNA, comp 2.8e-84 304 20475| 109492| 109492| M26232 Rat peripherin gene, complete cds. 1.4e-1 39 305 20472| 109006| 109006| S80636 injury-inducible gene (ID repetitive element, clon 3.6e-07 306 20471| 109001| 109001| M89646 Rattus norvegicus ribosomal protein S24 mRNA seque 1.6e-80 307 20468| 108618| 108618| L35240 Human enigma gene, complete cds. 2.5e-17 308 20467| 108615| 108615| X55572 R. norvegicus Apo D mRNA. 9.0e-193 309 20464| 108534| 108534| X77953 R. norvegicus mRNA for ribosomal protein S15a. 1.4e-68 310 20455| 106217| 106217| M20480 Mouse brain neurofilament-L mRNA, complete cds. 4.7e-122 311 20448| 106118| 106118| L10426 Mus musculus ets-related protein 81 (2R81) mRNA, c 5.1e-48 312 20443| 105946| 105946| D83407 ZAKI-4 mRNA in human skin fibroblast, complete cds 5.8e-63 313 20434| 105919| 105919| X61294 R. norvegicus L1 retroposon, ORF2 mRNA (partial). 1.5e-92 314 20430| 105905| 105905| D83783 Human mRNA for KIAA0192 gene, partial cds. 3.4e-05 315 20423| 105878| 105878| U13369 Human ribosomal DNA complete repeating unit. 0.021 316 20420| 105801| 105801| D17577 Mouse mRNA for kinesin-like protein (Kif1b), compl 2.5e-122 317 20419| 105787| 105787| X60370 R. norvegicus mRNA for microtubule associated prote 1.2e-97 318 20416| 105774| 105774| U02983 Rattus norvegicus secretogranin III (SgIII) mRNA, 9.0e-82 319 20412| 103932| 103932| M35826 Rat mitochondrial NADH-dehydrogenase (NDI) gene, c 2.0e-78 320 G protein signaling 20410| 103925| 103925| M12673 Rat guanine nucleotide-binding protein G-s, alpha 4.1e-66 321 20406| 103909| 103909| M13100 Rat long interspersed repetitive DNA sequence LINE 4.3e-172 322 20404| 103831| 103831| D90005 Rat endogenous retroviral sequence, 5′ and 3′ LTR. 4.8e-97 323 20403| 103830| 103830| M23953 Rat carboxypeptidase B gene, exons 6, 7, and 8. 1.1e-68 324 20401| 103821| 103821| D10699 Rat mRNA for ubiquitin carboxyl-terminal hydrolase 3.1e-99 325 20400| 103820| 103820| X60469 R. rattus FE65 mRNA for adaptor protein interacting 3.0e-167 326 20396| 103801| 103801| AF043698 Caenorhabditis elegans cosmid C54G6. 0.12 327 20395| 103795| 103795| X70369 R. norvegicus mRNA for pro alpha 1 collagen type II 3.0e-69 328 20391| 103703| 103703| V01556 Rat mitochondrial genome fragment from Sprague-Daw 1.0e-188 329 20385| 102872| 102872| L40632 Mus musculus epithelial ankyrin 3(7kb isoform) mR 1.6e-175 330 20379| 102748| 102748| M16597 Human cytochrome c1 mRNA, partial cds. 2.9e-127 331 20363| 101197| 101197| G05883 human STS WI-6610. 1.9e-65 332 20437| 101132| 101132| M89646 Rattus norvegicus ribosomal protein S24 mRNA seque 6.2e-55 333 hsp 20343| 101112| 101112| M86389 Rat heat shock protein (Hsp27) mRNA, complete cds. 1.2e-147 334 20342| 101108| 101108| X75683 H. spaiens (TL35) mRNA from LNCaP cell line. 1.0e-73 335 20340| 101083| 101083| X79064 V. unguicutata (Red Caloona) AKCS9 mRNA. 0.041 336 20334| 100940| 100940| Z12152 R. norvegicus mRNA for neurofilament protein middle 1.5e-206 337 20328| 100875| 100875| L01693 Rat peptidyiglycine alpha-amidating monooxygenase 3.2e-123 338 20322| 100839| 100839| X96487 B. taurus mRNA, anonymous. 5.5e-09 339 prteasome 20321| 100836| 100836| M29859 Rat proteasomes C2 component mRNA, complete cds. 1.0e-100 340 20317| 100759| 100759| M29341 Rat brain glyceraldehyde-3-phosphate dehydrogenase 2.8e-103 341 apoptosis 20308| 100704| 100704| D17614 Rat mRNA for 14-3-3 protein theta-subtype, complet 3.9e-228 342 20307| 100574| 100574| M22012 Mus musculus synaptosomal associated protein 25 (S 4.3e-72 343 20306| 100573| 100573| L27153 Mouse kinesin heavy chain (Khcs) mRNA, 5′ end. 1.0e-81 344 20302| 100557| 100557| U80954 Caenorhabditis elegans cosmid T07F8. 0.84 345 axon outgrowth 20301| 100465| 100465| X03369 Rat mRNA for beta-tubulin T beta15. 7.3e-91 346 20300| 100463| 100463| X62952 R. norvegicus mRNA for vimentin. 1.2e-114 347 20298| 100455| 100455| U62800 Human cystatin M (CST6) mRNA, complete cds. 8.9e-91 348 20296| 100438| 100438| A08267 Nucleotide sequence for P. falciparum RMA-1 gene. 0.87 349 20295| 100437| 100437| S69874 C-FABP=cutaneous fatty acid-binding protein [rats, 6.7e-160 350 20272| 1065569| Z33905| nicotinic acethyicholine receptor associated 43 kD protein 351 20264| 917438| Z26634| ankyrin brain variant, ANKB 352 20258| 1260365| Z21876| H. sapiens X-linked ALD gene. 353 GPCR 20238| 700123692| Y10148| NTR2 receptor 354 20204| 613665| X96427| Homol. to ScUBC9 355 20127| 921632| X69292| myosin heavy chain. Cell Type:smooth muscle Tissue Expression: GI wall, Respiration 356 20080| 1497875| X61615| H. sapiens mRNA for leukemia inhibitory factor (LIF) receptor 357 20055| UI-R-C1-kp-d-10-0-UI| X57748f properdin 358 20029| 700247749| X54673| H. sapiens GAT1 mRNA for GABA transporter 359 20027| 1244915| X54297| Human mRNA for histidine decarboxylase (EC 4.1.1.22) 360 19976| 700247354| X1S376| Human mRNA for GABA-A receptor, gamma 2 subunit 361 19960| 700247209| X14767| Human mRNA for GABA-A receptor, beta 1 subunit 362 transcription factor 19941| UI-R-BT0-pm-c-04-0-UI| X12740| C-Jun of the transcription factor AP-1 363 neuroptide 1986| UI-R-A0-au-d-10-0-UI| AA818532| Rat gamma-preprotachydinin A mRNA, complete cds 364 neuropeptide 19136| 315382| W12030| U10071 RNU10071 Rattus norvegicus CART protein mRNA, complete cds. Blastn: P. = 8.5e-117 365 proteasome 18970| 931025| U75362| ubiquitin carboxy terminal hydrolase 366 growth factor 18954| UI-R-Y0-mo-d-05-0-UI| U66198| fibroblast growth factor, FGF13 367 apoptosis 18941| 1050567| U59746| Bcl-w 368 18937| UI-R-AG0-xc-h-06-0-UI| U58522| huntingtin interacting protein 369 proteasome 18904| 407454| U44839| A ubiquitin C-terminal hydrolase 370 18871| 700128068| U33267| Human glycine receptor beta subunit (GLRB) mRNA, complete cds 371 18862| 803318| U30509| Gamma Glu Tranterase 372 18856| UI-R-C1-lp-e-07-0-UI| U28807| NFATc3 373 18835| 789552| U20240| CCAAT enhancer binding protein gamma 374 GPCR 18815| 700498633| U16129| Glutamate receptor (GluR4) 375 18784| 1617912| U08107| HSU08107 Human N-methyl-D-aspartate receptor subunit splice variant hNR1N (GRIN1) mRNA, partial cds. 376 1824| UI-R-A0-ar-f-11-0-UI| AA818840| Similar to MMU75839 mouse interferon regulatory factor 3 (mirf3) mRNA, alternatively spliced, complete cdsmRNA 377 18104| 1039340| M9S106| CREB-1 378 neuropeptide 18088| 640127| M88355| MUSOXYNEUI Mouse oxytocin-neurophysin I gene, complete cds. 379 18037| UI-R-C3-ss-b-09-0-UI| M737461 Homo sapiens lutropin/horiogonadotropin reeptor (LHCGR) mRNA, complete cds. 380 17418| UI-R-Y0-acb-c-12-0-UI| R60006| HUMEAR17A Human trilodothyronine recptor (THRAI, ear1), and ear2 genes, last 2 exons each. BLASTN P. 2.8E-96 381 1571| UI-R-A0-bc-d-07-0-UI| AA819345| Rat parvalbumin mRNA, complete cds 382 14951| 700861309| M59980| K Channel 383 14925| UI-R-C0-ie-h-11-0-UI| M55040| Human acetylcholinesterase (ACHE) mRNA, complete cds 384 14871| 492525| M29871| Human ras-related C3 botulinum toxin substrate (rac) mRNA, complete cds 385 G protein signaling 14870| UI-R-E0-da-b-11-0-UI| M29870| Human ras-related C3 botulinum toxin substrate (rac) mRNA, complete cds 386 14866| UI-R-AA0-wp-c-07-0-UI| M29548| Human elongation factor 1-alpha (EF1A) mRNA, partial cds. 387 14866| 299720| M29548| Human elongation factor 1-alpha (EF1A) mRNA, partial cds. 388 axon outgrowth 14839| 338087| M25667| Human neuronal growth protein 43 (GAP-43) mRNA, complete cds 389 G protein signaling 14829| UI-R-C1-lh-b-02-0-UI| M23612| Human GTPase-activating protein (GAP) mRNA, complete cds 390 14817| 349760| M21571| Human platelet-derived growth factor (PDGFA) A chain mRNA. 391 14739| 700124548| L76224| Homo sapiens NMDA receptor mRNA, complete cds 392 14727| 1162580| L42198| MUSALOX Mus musculus arachidonate 5-lipoxygenase (Alox5) gene, complete cds. 393 14677| 1066814| L16510| Homo sapiens cathepsin B A cysteine protease associated with tumor invasiveness and neovascurlarisati 394 GPCR 1467| UI-R-A0-an-d-02-0-UI| AA819622| Rat G-protein coupled thrombin receptor mRNA, complete cds 395 neuropeptide 14620| 482891| K01911| Human neuropeptide Y (NPY) mRNA, complete cds 396 13845| 963216| H58254| monocyte chemoattractant protein 1 receptor (MCP-1RA) 397 13310| 602816| D86323| D86323 Mus musculus mRNA for calmegin, complete cds. 398 13304| 1246629| D83004| similar to Drosophila bendless gene 399 proteasome 13275| UI-R-E0-ct-g-08-0-UI| D23662| ubiquitin-like protein 400 13165| UI-R-BT0-qi-f-04-0-UI| AI145577| Rat degenerin channel MDEG mRNA, complete cds 401 hsp 1278| 1246430| AA823192| D49547 HUMHSP40 Human mRNA for heat-shock protein 40, complete cds. Blastn P. = 3.5E-167 402 12712| UI-R-C2-nf-c-01-0-UI| AI072073| Rat vasopressin mRNA, complete cds 403 G protein signaling 12702| UI-R-C2-nh-g-02-0-UI| A1071860| Rat regulator of G-protein signalling 14 (RGS14) mRNA, complete cds 404 12681| UI-R-C2-my-e-04-0-UI| A1071179| Rat mRNA for cyclinG-associated kinase, complete cds 405 12678| UI-R-C2-ms-b-07-0-UI| AI071095| Rat vgr mRNA 406 1266| UI-R-AO-aq-c-10-0-UI| AA819897| Rat synuclein 1 mRNA, complete cds 407 ion channel 12538| UI-R-C1-kp-b-07-0-UI| AI058645| Rat Kv4.3 potassium channel mRNA, complete cds 408 neuropeptide 12490| UI-R-C1-iz-a-09-0-UI| AI045437| Rat neuropeptide Y mRNA, complete cds 409 12463| UI-R-C1-kd-e-12-0-UI| AI044870| R. norvegicus mRNA for elongation factor 1 alpha 410 12448| UI-R-C1-kc-c-05-0-UI| AI044610| Rat dopa decarboxylase (DOC) mRNA, complete cds 411 12427| UI-R-C1-kb-b-03-0-UI| AI044262| Rat mRNA for thyrotropin releasing hormone receptor, complete cds 412 12350| UI-R-C0-ib-c-02-0-UI| AI030286| Rat brain-derived neurotrophic factor (BDNF) mRNA, complete cds 413 12318| UI-R-CO-iy-b-07-0-UI| AI029599| Rat mineralocorticoid receptor (MR) mRNA, complete cds 414 12263| 659316| AF075599| conjugating enzymes 415 ion channel 12260| 700254638| AF064876| BCNG-1 hyperpolarization NSC channel=HCN1 = HAC2 416 proteasome 12223| UI-R-C1-kt-f-03-0-UI| AF022789| ubiquitin hydrolyzing enzyme I 417 11788| UI-R-C0-ho-b-04-0-UI| AA997314| Rat cDNA clone UI-R-C0-ho-b-04-0-UI 3′ mRNA 418 proteasome 1038| UI-R-E0-cj-b-04-0-UI| AA859300| Rat proteasomes C2 component mRNA, complete cds 419 proteasome 1031| UI-R-E0-cd-c-03-0-UI| AA859229| Rattus norvegicus ubiquitin conjugating enzyme mRNA, complete cds C D E F G H I J K 1 CA1.CMS CA1.IMP CA1.IMPnCMS CA1.JRF CA1.JRFnCMS CA3.CMS CA3.IMP CA3.IMPnCMS CA3.JRF 2 1 1 1 1 1 1 1 1 1 3 1 1 1 1 1.135106383 1 1 1 1 4 1 1.026595745 1.069148936 1.109148936 1.159787234 1.053803876 1.536399576 1.540545753 1.389933468 5 1.283765785 1.117871942 1.152476322 1.417151736 1.500295975 1.098971902 0.830980194 0.88179679 0.923074768 6 0.985121588 1.016195517 1.146686505 1.191468085 1.299080465 1.229552015 1.146405788 0.918101144 1.264264585 7 1 1 1 1 1.154787234 1 1 1 1 8 0.918237765 1.515092312 1.8366709 2.101396894 1.508840481 0.833539644 0.862206007 0.926900775 0.864616587 9 1.271983351 1.490840538 1.058110556 1.17292499 1.134983235 1.184266024 0.98016667 0.86311757 1.142719482 10 0.786262347 0.879611818 1.108889705 1.083839454 1.490824994 0.867116391 1.20458322 1.08546415 1.020460802 11 1.277605055 1.843345703 1.294554207 1.527329255 1.789088356 1.071466291 0.968051399 0.937863052 1.243531068 12 0.580658633 0.857415583 0.803006801 0.723660661 0.930258919 0.927113703 0.854713314 1.140751539 1.097910593 13 0.760842082 0.967954708 1.158522509 0.800087946 0.68949596 0.778530809 0.868427203 0.784918595 0.732258316 14 1 1.955319149 1.436808511 1.609787234 1.482234043 1 1.249255311 1.294361702 1.601808511 15 1 1 1 1 1 1 1 1 1 16 1 1 1.083085106 1 1 1 1 1 1 17 1.68533171 1.843919659 1.306025563 1.025562995 0.761412051 1.042526807 0.870960683 1.147676709 0.991100737 18 0.918452657 1.022825294 1.167341251 1.187777852 1.200384845 1.010731303 1.033229741 0.934244873 0.893184172 19 1.031509395 0.932438571 1.124055338 0.858145777 1.115713401 0.971141137 0.83235423 0.850822441 0.850712293 20 1 1 1 1 1 1 1 1 1 21 1.330920372 1.107606572 1.370217166 1.141388027 1.355883029 1.197396918 1.041212021 22 1.001916525 1.729840999 1.175042589 1.122373651 1.216283362 0.8854339 1.01981501 1.018987818 0.797337946 23 1.168235333 0.868461614 0.829895386 0.880070836 0.817892631 1.282736246 1.151929311 1.217781459 0.648857596 24 0.806988969 1.21877545 1.287776708 1.377952173 1.213148738 0.896036998 0.983004963 1.312265002 0.995262445 25 1.548089532 1.22034818 1.203527018 1.708161881 1.663124576 1.102383239 1.001680874 1.09205787 1.264227398 26 1 1.348191489 1.561276596 1.589361702 1.994787234 0.859468723 0.972812963 0.773545221 0.775103217 27 1.128055186 0.937950837 0.853841305 0.756985783 0.825232859 0.972462265 0.848513654 0.910617583 0.784796636 28 1 1.147340426 1.153297872 1.194255319 1.194361702 1 1 1 1 29 0.929867332 1.0147409 1.260800544 1.333484522 1.522678308 0.83777447 0.778861183 0.844324525 0.689691105 30 1 1 1 1.025 1 1 1 1 1 31 1.113756425 0.948850657 1.197387207 1.197208738 1.221159338 1.064588104 1.141800247 0.889935999 0.918677705 32 1.001651053 0.972291016 0.96317433 1.059976311 0.713398658 0.927060922 1.004460303 0.816130556 0.828514457 33 0.915764112 0.499164687 0.602538896 0.708686116 0.673568888 0.884301786 1.584374032 1.231258816 1.503423126 34 0.707739249 1.05150544 1.265663679 1.446508415 1.305367389 0.958500813 0.900369795 1.187684227 0.844704075 35 1.045635922 1.026555979 1.459111131 1.488602486 1.335229097 0.89865223 1.152229878 1.020670856 1.020784432 36 0.908402292 1.832463399 1.196721833 1.174633991 1.095798854 1.068026865 1.253434415 1.775770835 1.577592348 37 1.113297872 1.338085106 1.106489362 1.1752127661 0.853193351 1.20962379 1.209623797 1.035053952 0.86573345 38 0.942528893 0.933867927 0.855850933 1.025504768 0.825086746 1.132523583 1.276845591 1.315607869 0.995005148 39 1 1.016489362 1.344042553 1.98 1.780212766 1 1 1 1 40 1 1.036595745 1 1 1 1 1.059574468 1 1 41 1 1.036595745 1 1 1 1.030212766 1.249255319 1.164361702 1.174574468 42 0.928423354 0.98207757 1.330933625 1.296577588 1.274000226 0.982149165 1.018675082 0.981466218 0.847396557 43 0.971175085 1.263510529 1.663230747 2.063944927 1.958732145 1.007528423 0.938664374 1.229750645 0.800496799 44 1.205319149 1.418404255 1.882234043 1.958297872 1.946170213 1.118771009 1.02866073 1.150380374 1.142772896 45 1 1 1 1 1 1 1 1 1 46 1.694640812 1.304332372 0.958868133 1.026420757 0.89092058 1.539610003 0.876365457 0.585746033 0.952971924 47 0.806988969 0.966165692 0.875101799 0.891241578 0.886651366 1 1 1 1 48 0.808675141 0.578299043 0.655571572 0.466998646 0.66823921 1.209680851 1.279148936 1.128297872 49 1.00393617 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0.944476918 2.242061507 341 0.80786679 0.953809671 0.880296938 1.032677978 0.974055122 1.360220846 0.972035192 0.996139689 2.539557286 342 1.198477009 0.846919848 0.758322757 1.246484086 1.034830167 1.184513529 1.224401678 1.06591825 2.224170182 343 1.460210155 0.806304651 0.713718863 1 1 1 1 1 1.557351753 344 0.964386792 0.898820755 0.915978774 0.959186702 1.283689522 1.012540813 1.105224102 1.166295637 2.29122733 345 0.7148567 0.844718197 0.910898504 1.463194311 1.488966156 1.477993465 1.362780214 1.561931054 2.115540522 346 1.909893617 1.057872341 1.085212766 1.102978723 1 1.198297872 1 1.909893617 347 1.003190868 0.880814345 1.072356299 0.897957921 0.803960396 0.825618812 0.703155941 0.639665842 1.847992236 348 1.066872149 0.97531747 1.190062878 0.987956013 1.133317818 0.977075697 0.858943775 0.980954597 1.974879917 349 1.118065108 0.924438331 1.097294819 0.985637657 1.022469554 0.957472088 0.817535338 1.041933849 1.985113083 350 1.387174997 1.092738785 1.083093089 0.91083237 0.979242653 1.084304481 0.998754559 1.201405569 1.8 351 1.709242784 1.193580685 1.002460165 0.932237085 0.816364204 0.380335691 1.011416887 0.949504416 2.261063261 352 1 1 1 1 1.28606383 1.143297872 1 1 1.503085106 1.503085106 353 1 1 1.199787234 1.197776806 0.947542622 1.208767876 1.128395679 0.808030975 1.943297872 354 1.135135135 0.98540238 0.905960065 1.015951779 0.87506469 0.607385309 0.788304058 0.663429633 1.436937879 355 0.686164595 1.03285633 0.903833432 0.648813898 1.16167316 0.875455793 0.981213693 1.072663496 1.201913442 356 1 1 1 1 1 1 1 1 1.867021277 357 0.88360404 1.201796435 1.075254805 0.634219197 0.743879531 0.66615342 0.729713916 0.769008099 1.618989618 358 1 1 1 0.984644592 1.282818695 1.355910436 1.389804009 1.30135686 2.656492544 359 1 1 1 1 1 1 1 1 1.859574468 360 0.833154982 1.110526799 0.960834072 1.080841669 1.038022296 1.142849355 1.086974297 1.115211644 1.357630358 361 0.864465799 0.944717378 0.951492739 1.46970117 1.221017908 1.399318068 1.17026113 1.216443976 3.190718592 362 0.743494424 1.008700467 0.858024203 1.127217125 1.267787971 1.164424057 1.117940877 1.142813456 1.267787971 363 1 1 1 1 1 1 1 1 3.235425532 364 0.827611891 0.863374614 0.802143318 0.903736627 0.74858649 0.863259083 1.044384723 0.665874205 1.791358517 365 1.071489362 1.099042553 1.042659574 1.18287234 1.031914894 1.06 1.198297872 1.015531915 1.7483877097 366 1 1 1 1.020552344 1.25613359 1.433590238 1.15433526 1.573281952 2.259707193 367 0.711997477 0.78493178 0.768001195 1.040069409 0.90072593 1.357472618 1.191129648 1.173522669 2.114286534 368 1 1 1 1 1 1 1 1 2.150044085 369 0.759793671 0.938630977 0.862456434 0.972080028 0.994004038 1.205127159 1.191034609 1.072910081 4.27175051 370 1.57453322 1.356098643 1.302717492 1.062281282 1.093525553 1.24456942 1 .047538004 0.932242302 2.595615003 371 1.072189728 1.227946479 1.205352135 0.766533475 0.766533475 0.816602789 0.910462366 0.892848406 1.278343081 372 1.779403382 1.459908797 1.645259358 0.682563714 0.89822525 0.588695433 0.671124569 0.558541509 1.779403382 373 0.716081359 0.942637312 0.846042508 1.069335239 1.57692995 1.196568978 1.443084346 1.587651894 2.166276596 374 1.199953293 1.059831854 0.978701541 1.103592248 0.861430597 0.96608634 1.16925319 1.033480384 1.929473433 375 1.169293409 0.896377979 1.199151752 1.007031598 1.171384068 0.774032043 0.612527815 0.983622608 2.234498475 376 1.04996117 1.056777979 1.212787989 0.943159923 0.986062942 0.801734104 0.767951188 0.831021195 1.93501115 377 1.080735412 1.119765058 0.957877177 0.743297566 0.867850473 0.675339474 0.688380337 0.70999905 1.915590267 378 0.94493137 0.777034803 0.912809884 0.641521117 0.4714333 0.355468583 0.252481648 0.566878906 1 379 1 1 1 1 1 1 1 1 2.194099834 380 0.82255303 0.891229906 0.925853705 0.976414891 1.006929202 1.028233269 0.867398321 0.778738971 1.934825035 381 1 1 1 1 1 1 1 6.955212766 382 0.773165896 0.848446794 0.835228024 0.936721728 0.862092334 1.237018213 0.827869547 0.796272766 1.580106383 383 0.687036906 0.976642439 0.844047703 0.875885203 1.176108833 1.069139023 0.984811778 1.173686172 3.240602166 384 1 1 1 1 1.123191489 1 1 1.303404255 1.433004158 385 0.737497635 0.877719619 0.887368355 1.15385234 1.036244705 1.34389577 1.147954533 1.191491073 2.005849966 386 0.646593945 1.055209261 0.968239834 0.957858255 1.306042493 1.041890602 1.05952082 1.098648852 1.829488465 387 1.021246344 1.102353013 0.99348459 1.078375967 1.103036612 0.918838895 1.565018117 1.341280893 1.565592001 388 0.88612368 0.88612368 0.88612368 0.834412436 0.630571455 1.146757999 0.897657252 0.763723374 3.199516212 389 0.952863659 0.952863659 0.952863659 1 1.193617021 1 1 1.052978723 2.160636183 390 1.061572453 0.969276305 0.937760547 0.942207265 0.884240455 1.023151909 0.938725775 0.912556574 1.964051968 391 1 1 1 1 1 1 1 1 1.709526136 392 0.627583943 0.962766272 1.011369679 1.103367342 1.21882135 1.142357618 1.18998579 1.209433028 1.301790471 393 1.427132701 1.188684834 1.250592417 0.748587363 0.966903941 1.237543113 1.192338739 1.681147721 1.681147721 394 2.359800149 1.718992641 1.529066235 0.929832768 0.905430164 0.608505828 0.756207851 0.777764784 2.359800149 395 1 1 1 1 1 1 1 1 2.042765957 396 1 1 1 1 1 1 1 1 2.3688923 397 1.354947775 0.922840065 0.853589326 1.250662589 1.191641182 1.092558614 1.208766565 1.124974516 1.581131544 398 1.040114326 1.35270521 1.124103696 1.391611118 1.637274393 1.378513234 1.591743656 1.673059681 1.673059681 399 0.820279246 0.748987271 0.761345644 0.936766312 1.288493038 1.043987169 1.00996451 1.12465875 2.046190397 400 0.937188435 0.937188435 0.937188435 1.014633836 1.03906598 1.291467837 1.082712986 1.120188331 1.954148936 410 1 1 1 1 1 1 1 1.090319149 2.033727592 402 1 0.926659063 0.973127939 0.872309105 0.697975742 0.686815505 0.464098545 0.717216693 1 403 1 1 1 1 1 1 1 1 2.635141623 404 1.079420019 0.994480823 1.134705332 1 1.011595745 1.129468085 1 1.275851064 1.849148936 405 1.261271141 1.096042306 1.294779939 1.106045922 1.001480202 1.201325133 1.221889372 1.006061781 1.481142714 406 0.769099277 0.838461538 0.841880342 0.958205912 1.162895005 0.958205912 0.965749235 1.089398573 1.805638298 407 1 1 1 1 1.011595745 1 1 1.043617021 1.238758251 408 1 1 1 1 1 1 1 1 3.180443147 409 0.866921683 0.859707152 0.85909311 1.11047861 0.951699463 1.11047861 1.11047861 1.01601536 1.111936029 410 1 1 1.052446809 2.281276596 1.868404255 1.759148936 1.583510638 2.281276596 411 1.320499623 1.093816485 1.292636058 1.098297872 1 1.011914894 1 1.006170213 1.868606199 412 1 1 1 1 1 1 1 1 1.07477641 413 1 1 1 1 1 1 1 1 1.816303024 414 0.843774481 0.887145401 0.880545994 1.18847045 1.107964173 1.324549573 1.310059826 1.22198015 2.134586036 415 1.162773534 1.09509576 1.006310196 0.736155759 0.765694183 0.742342452 0.8100928 0.626978832 2.034803032 416 0.679366408 0.971530249 0.97494941 1.076604118 0.938927542 1.08995713 1.074003795 1.022559561 2.041276596 417 0.591414614 0.878754807 0.853861345 1.151216609 1.097328749 1.400952129 1.163118223 1.104601957 2.185061528 418 0.769526277 0.848212092 0.979920934 1.211142695 0.867879477 1.227743036 1.042455941 1.074792496 2.999187747 419 0.842346904 1.10073594 1.092836405 1.282111488 1.065310027 1.094356031 1.18505957 1.323419428 1.680040323 AM AM AM AM 1 min 1 min 1 min 1 min 2 0.617720674 49 0.27783982 96 0.657987256 143 0.602525479 3 0.975882179 50 0.602448247 97 0.382084802 144 0.604985472 4 0.374159137 51 0.384386423 98 0.338017625 145 0.461479624 5 0.434105083 52 0.451640801 99 0.893026791 146 0.57766974 6 0.499975309 53 0.366885023 100 1 147 0.49659253 7 0.493257071 54 0.757270603 101 0.638500204 148 1 8 0.643130816 55 0.712224067 102 0.679807998 149 0.189006771 9 0.750773905 56 0.872632752 103 0.747921832 150 0.54598338 10 0.395613834 57 0.79459618 104 0.685190305 151 0.936161737 11 0.770591549 58 0.6006124 105 0.477289307 152 0.562493219 12 0.580658633 59 0.855115112 106 0.813003946 153 1 13 0.68949596 60 0.669166917 107 0.558380572 154 0.129959906 14 0.80129047 61 0.449564477 108 0.677042676 155 0.640803312 15 0.720711954 62 0.576620499 109 0.470047005 156 0.609360819 16 1 63 0.48107104 110 0.647026432 157 0.522579204 17 0.699456805 64 0.312010272 111 0.685770313 158 0.362416767 18 0.716511794 65 0.253919966 112 0.324518401 159 0.46555396 19 0.692804106 66 0.602417457 113 0.305283129 160 0.648650284 20 0.501467058 67 0.776800864 114 0.412230029 161 0.336123336 21 0.65582404 68 0.497450765 115 0.287437055 162 0.433025559 22 0.534945144 69 0.496881277 116 0.547455016 163 0.562022224 23 0.394132914 70 0.748288489 117 0.434842948 164 1 24 0.46631031171 71 0.365505203 118 0.387965671 165 0.861169898 25 0.448302175 72 0.399300334 119 0.49564269 166 0.545835251 26 0.773545221 73 0.395935179 120 0.767008097 167 0.429089161 27 0.736030759 74 0.756478352 121 1 168 0.863502705 28 0.551247861 75 0.670402612 122 0.753109815 169 0.390472833 29 0.689691105 76 0.670934199 123 0.401368018 170 0.744555027 30 1 77 0.743553235 124 1 171 0.897061304 31 0.529070749 78 0.781681497 125 0.384505256 172 0.988002181 32 0.52713633 79 0.751830581 126 0.474651586 173 0.520525452 33 0.228436886 80 0.499550979 127 0.215310085 174 0.447567811 34 0.669884546 81 0.515803336 128 0.723447772 175 0.877684407 35 0.320223641 82 1 129 0.963806008 176 0.669475875 36 0.894378651 83 0.707405178 130 0.677427259 177 0.419926014 37 0.555065854 84 0.587066076 131 0.309265271 178 0.735483302 38 0.358184612 85 0.293988866 132 0.558485656 179 0.378208739 39 1 86 0.591317099 133 0.370794052 180 0.494470842 40 0.53972894 87 0.723738504 134 0.214881701 181 0.450794404 41 0.716081359 88 0.404818059 135 0.826594261 182 0.220388258 42 0.503253302 89 0.5147789 136 0.446090585 183 0.540766551 43 0.55864057 90 0.556552798 137 0.485863441 184 0.798910963 44 0.709172555 91 0.755960401 138 0.515803336 185 0.425252618 45 0.930600931 92 0.700735655 139 0.654130134 186 0.864769686 46 0.585746033 93 0.786977492 140 0.266284397 187 0.529378221 47 0.806988969 94 0.799574065 141 0.4427342 188 0.29353339 48 0.194584748 95 0.693984525 142 0.444107358 189 0.35295324 AM AM AM AM AM 1 min 1 min 1 min 1 min 1 min 190 0.552892751 237 0.758157046 284 0.597972935 331 0.511926805 378 0.252481648 191 0.509159097 238 0.402978098 285 0.452363537 332 0.378309811 379 1 192 1 239 0.606356833 286 0.648215892 333 0.332484437 380 0.587932629 193 0.308267346 240 0.556938026 287 0.442389849 334 0.467057538 381 0.172698879 194 0.884632717 241 0.838807581 288 0.69163417 335 0.459163736 382 0.445683898 195 0.551287315 242 0.335910908 289 0.621766437 336 0.577861844 383 0.506510235 196 0.726254105 243 0.15105012 290 0.517398509 337 0.46292613 384 0.344836285 197 0.42560609 244 0.557855078 291 0.546225767 338 0.610106902 385 0.626518866 198 0.533936202 245 0.727936174 292 0.680963117 339 0.422928102 386 0.440755849 199 0.62401015 246 0.907906227 293 0.580263483 340 0.611884939 387 0.433022751 200 0.558559629 247 0.804729056 294 0.738023298 341 0.689945904 388 0.630571455 201 0.756478352 248 0.721080086 295 0.489583333 342 0.687276256 389 0.630957175 202 0.175626909 249 0.814464609 296 0.551675568 343 0.44078182 390 0.884240455 203 0.506079468 250 0.631698186 297 0.73151751 344 0.621230121 391 0.459208598 204 0.553722903 251 0.548784249 298 0.82673197 345 0.627326099 392 0.544781894 205 0.336076818 252 0.715100799 299 0.594965599 346 0.327937483 393 0.45538223 206 0.665387843 253 0.680674967 300 0.419221373 347 0.624469247 394 0.480167057 207 0.236429103 254 0.735996688 301 0.472338073 348 0.728834893 395 1 208 0.624969 255 0.525051826 302 0.738377393 349 0.817535338 396 1 209 0.683054298 256 0.497034721 303 0.648374627 350 0.645352892 397 0.516248974 210 0.390673704 257 0.717037778 304 0.532282899 351 0.358372728 398 0.549846837 211 0.741461145 258 0.202296504 305 0.444067596 352 0.304907717 399 0.65972613 212 0.557173245 259 0.436500972 306 0.315068933 353 0.75365 400 0.666051979 213 0.598293009 260 0.381803575 307 0.718215812 354 0.359324957 401 0.915109034 214 0.456645908 261 0.340460584 308 0.593494116 355 0.393581046 402 0.464098545 215 0.607909148 262 0.796473838 309 0.509321345 356 1 403 1 216 0.504449319 263 0.4361085 310 0.590813094 357 0.528711619 404 0.629888348 217 0.410367452 264 0.863172582 311 0.587056544 358 0.487956811 405 0.33895512 218 0.425422724 265 0.687687688 312 0.474594392 359 1 406 0.489437371 219 0.373219281 266 0.681356915 313 0.22932982 360 0.499916185 407 0.385313531 220 0.613000421 267 0.784313725 314 0.508520422 361 0.812983789 408 1 221 0.234759372 268 0.53946088 315 0.416349019 362 0.541568008 409 0.493831628 222 0.563481 597 269 0.348793238 316 0.586489527 363 1 0.52660431 410 0.560583312 223 0.404874069 270 0.942544871 317 0.451528634 364 0.52660431 411 0.680095244 224 1 271 0.080946558 318 0.62767094 365 0.543667149 412 0.538219296 225 0.324238557 272 0.382358758 319 0.496851701 366 0.441336067 413 0.8358868 226 0.212409869 273 0.658920874 320 0.163060943 367 0.489400859 414 0.754236235 227 0.210617132 274 0.541104912 321 0.101068748 368 1 415 0.626978832 228 0.547898493 275 0.123670821 322 0.475236278 369 0.713372638 416 0.679366408 229 0.791778976 276 0.782523969 323 0.525896759 370 0.582719279 417 0.591414614 230 0.634663426 277 0.625202102 324 0.682514738 371 0.347890452 418 0.644690782 231 0.378419422 278 0.527003795 325 0.347704715 372 0.44009551 419 0.550725293 232 0.600271657 279 0.58435907 326 0.415432023 373 0.388333471 233 0.884189508 280 0.862417805 327 0.468477448 374 0.272697033 234 0.44825685 281 0.681634155 328 0.484884222 375 0.406168567 235 0.582589908 282 0.754535238 329 0.802321611 376 0.767951188 236 0.550187401 283 0.381947134 330 0.7546261 377 0.625584235 

What is claimed is:
 1. A polynucleotide microarray comprising at least one polynucleotide set forth in Table 1, Table 2, Table 3 or Table 4, wherein expression of said polynucleotide is either increased or decreased in brain cells in response to stress compared to normal brain cells.
 2. A method for screening a compound for effectiveness in altering expression of a target polynucleotide wherein said target polynucleotide comprises a polynucleotide selected from Table 1, Table 2, Table 3 or Table 4, wherein said polynucleotide expression is either increased or decreased in brain cells in response to stress compared to normal brain cells.
 3. A method of treating depression in a mammal comprising administering a compound identified in the method of claim
 2. 